Bioengineered Livers in the
Classroom, You can’t
Really do that?
Robb Bartenslager
Palm Beach Central High School
1
Table of Contents
Introduction………………………………………………………………………………………………………………..4
Lesson Summaries………………………………………………………………………………………………………5
Lesson Sequencing Guide………………………………………………………………………………….……….6
Lesson Vocabulary……………………………………………………………………………………....……………7-10
Next Generation Sunshine State Standards – NGSSS……………………………………………….11-12
Background Information…………………………………………………………………………………………….13
Unit Pre-Test…………………………………………………………………………………………………………….14-16
Unit Pre-Test Key…………………………………………………………………………………….…………………17
Lesson One “What is Regenerative Medicine?”………………………………………………………..18-32
Lesson One Teacher Pages………………………………………………………………………………………..18-27
Lesson One Student Pages…………………………………………………………………..……………………28-32
Lesson Two “What is a Liver?”…………………………………………………………………………………..33-38
Lesson Two Teacher Pages……………………………………………………………………..…………………33-36
Lesson Two Student Pages………………………………………………………..……………………………….37-38
Lesson Three “Diagnosing Diabetes”………………………………………………………………………....39-41
Lesson Four “Reading a Scientific Journal”…………………………………..……………………………..42-56
Lesson Four Teacher Pages…………………………………………………………………..……………………..42-50
Lesson Four Student Pages………………………………………………………………..…………………………51-56
2
Lesson Five “Bioengineer Me a Liver!: Part One”……………………………………………………….…57-70
Lesson Five Teacher Pages…………………………………………………………………………………………….57-66
Lesson Five Student Pages……………………………………………………………………………………………..67-70
Lesson Six “We Need Stem Cells”…………………………………………………………………………………..71-73
Lesson Seven “Bioengineer Me a Liver!: Part Two”…………………………………………………………74-79
Unit Post Test……………………………………………………………………………………………………………….…80-82
Unit Post Test Key……………………………………………………………………………………………………………..83
3
Introduction
In the summer of 2013, I had the opportunity to work in the Petersen Lab at the
University of Florida, under the direction of Dr. Bryon Petersen. This opened my eyes to
the relatively new branch of medicine known as regenerative medicine. Instead of
coming up with a new drug to treat symptoms, regenerative medicine has the goal of
replacing damaged organs with new organs bioengineered in the lab. Sounds like
science fiction? There have been many advances in the field over the last few years,
and students will get to see some of these through video clips produced by the
researchers.
The idea is to take an organ like a liver (the focus of the Petersen Lab) and
decellularize it. Various detergents remove all the cells, leaving only the extracellular
matrix (ECM) behind. This ECM is mostly collagen fibers, but is also thought to contain
some markers to stimulate tissue growth. Stem cells, either embryonic or induced
pluripotent stem cells, can then be seeded on the ECM scaffold to produce a new,
functioning liver. This has not been fully accomplished yet.
My task was to take my experience and develop a curriculum unit for high school
students. When the organs are removed from pigs, the pigs are first heparinized so the
blood will not clot in the vascular tissue. Researchers want to keep the vasculature
intact as it will provide a pathway for oxygen and nutrients for the new cells. Tubing is
attached to the hepatic artery and portal vein. A peristaltic pump is used to send a
detergent like Triton X 100 and SDS through the organ over a 1-2 day period to cause
cell lysis. How can you do this in your classroom? I developed a unique, low cost
method of using chicken livers available from your local grocer. You won’t be
decellularizing entire organs, but the result is pretty dynamic. Students will be amazed
when they see they have accomplished the first step of organ bioengineering!
4
Lesson Summaries
Unit Pre-Test
Lesson One: “What is Regenerative Medicine?”



A web quest introduces students to the field of regenerative medicine, focusing
on video clips from researchers.
A graph is produced using data from the CDC.
Percent increase is calculated using data from the CDC.
Lesson Two: “What is a Liver?”


Powerpoint presentation on liver anatomy, function and disease with
accompanying student note page.
Jigsaw activity on types of diabetes.
Lesson Three: “Diagnosing Diabetes”

Diabetes detection simulation from Science Takeout.
Lesson Four: “Reading a Scientific Journal Article”

High level reading about an experiment similar to the one students will be
conduction.
Lesson Five: “Bioengineer Me a Liver!: Part One”


Day one of liver decellularization
Reinforces the scientific method/experimental design.
Lesson Six: “We Need Stem Cells”

Stem cell simulation from Science Takeout.
Lesson Seven: “Bioengineer Me a Liver!: Part Two”


Day two of liver decellularization.
Analysis of results.
Unit Post-Test
5
Lesson Sequencing Guide
Week One
Day One
Day Two
Day Three
Day Four
Day Five
Unit Pre-Test
Lesson One
Lesson Two
Lesson Three
Lesson Four
Week Two
Day One
Day Two
Day Three
Day Four
Lesson Five
Lesson Six
Lesson Seven
Unit Post Test
6
Unit Vocabulary
Adult stem cell: Stem cells taken from umbilical cord blood, bone marrow, or some organs.
Allograft gold standard: An organ transplant between members of the same species, like from
one person to another.
Anesthetized: To be placed under anesthesia, or to lose sensation. Performed before surgery
so the patient won’t feel pain.
Apoptotic: When cells die off.
Artery: Blood vessel that carries oxygen rich blood away from the heart.
Bile: Fluid produced by the liver and stored in the gall bladder, breaks down fats.
Bioengineering: Using biology, chemistry and physics to come up with real world applications
for biological problems.
Bioprinting: The use of 3D printers to produce molds using biological materials like collagen.
Cannulated: Placing a small tube (cannula) into the body.
CDC: The U.S. Centers for Disease Control and Prevention. Part of the U.S. federal government,
the CDC tracks all things regarding public health, from viral outbreaks to obesity.
Cirrhosis: After chronic liver disease, fibrous tissue replaces liver tissue and liver loses function.
Constants: Factors that are kept the same throughout the experiment.
Control: Group that is not treated with the independent variable. Used to measure results
against.
Cytokine: Small protein molecules that are associated with inflammation.
Decellularization: The process of treating organs with detergents to remove all the cells,
leaving behind the extracellular matrix (ECM).
Dependent Variable: The change you are looking for, what you are measuring.
7
Diabetes: Disorder in which either the body makes no insulin (Type 1) or cells aren’t able to use
the insulin properly (Type 2).
Differentiation: Process an unspecialized cell takes to become specialized, changes form and
function.
Embryonic stem cell: Stem cell taken from an embryo.
Euthanized: Intentionally killing an organism.
Experimental Group: Group that is treated with the independent variable.
Extracellular Matrix (ECM): The non-living material that is between the cells that gives the
organ its shape, consisting mostly of collagen. Sometimes referred to as a scaffold for
bioengineering organs.
Fibrosis: Fibrous tissue replaces liver tissue, leads to cirrhosis.
Glucose: A monosaccharide or simple sugar (C6H12O6), it is the major fuel for cellular
respiration.
Growth factor: Proteins that turn genes on /off to promote specialization.
Heparinized: Treated with an anticoagulant (heparin) to prevent blood clotting.
Hepatic: Refers to the liver.
Hepatitis: General term for inflammation of the liver, can be caused by several pathogens.
Hepatocyte: Main type of cell found in the liver. Not highly specialized, can perform most liver
functions.
Histologically: Refers to the study of tissues.
Homeostasis: Maintaining a balance, or regulating the environment in the cells or body as a
whole.
Hormone: A chemical released by a cell, gland or organ and affects another part of the body.
Hypothesis: Prediction for the results of the experiment, best written as an “if-then” statement
and based on something measureable.
Immunogenicity: The ability of a substance to cause an immune response.
8
In vitro fertilization (IVF): Combining sex cells (sperm/egg) in the laboratory to create
embryos.
Induced pluripotent stem cell: An adult cell that has undergone a process to turn it into a stem
cell.
Insulin: A hormone made in the pancreas that allows cells to absorb glucose.
Kuppfer cell: A macrophage of the liver, it can clear out debris, old red blood cells and
pathogens.
Lobule: Small functional unit of the liver where blood flows into the sinusoids or spaces
between the blood vessels.
Lysis: Breaking apart a cell.
Macrophage: Large cells that perform phagocytosis, or ‘eating’ of foreign particles.
Pancreas: An endocrine/digestive gland that produces pancreatic juice and insulin.
Perfused: To permeate an organ with a fluid.
Porcine: Refers to a pig.
Proinflammatory: Promotes inflammation.
Protocol: A set of procedures or steps to be followed.
Receptor: A molecule on the cell membrane that receives chemical signals that direct the cell.
Regenerate: The ability to regrow and function. The liver is the only internal organ that can
regrow.
Regenerative Medicine: Branch of medicine that seeks to replace defective body parts with
new bioengineered parts.
Scaffold: The non-living architectural component of an organ, consisting primarily of ECM.
SDS: Sodium dodecyl sulfate, a detergent used to cause cell lysis.
Sham Operated: A fake or false operation. Used as a control in the context of this paper.
Stem Cell: A cell that can differentiate/change into many different types of cells.
9
Therapeutic: Has the ability to heal, reduce symptoms.
Tonal: Refers to different shade of color in the context of this paper.
Triton X 100: A non-ionic detergent used to cause cell lysis.
Vein: Blood vessel that carries oxygen poor blood towards the heart.
Xenogenic: From different species, as in a transplant from a pig into a rat.
Zygote: The first stage of development when the sperm joins the egg.
10
Next Generation Sunshine State Standards NGSSS
SC.912.N.1.1 Define a problem based on a specific body of knowledge, for example: biology,
chemistry, physics, and earth/space science, and do the following:
1. pose questions about the natural world,
2. conduct systematic observations,
3. examine books and other sources of information to see what is already
known,
4. review what is known in light of empirical evidence,
5. plan investigations,
6. use tools to gather, analyze, and interpret data (this includes the use of
measurement in metric and other systems, and also the generation and
interpretation of graphical representations of data, including data tables
and graphs),
7. pose answers, explanations, or descriptions of events,
8. generate explanations that explicate or describe natural phenomena
(inferences),
9. use appropriate evidence and reasoning to justify these explanations to
others,
10. communicate results of scientific investigations, and
11. evaluate the merits of the explanations produced by others.
(Lessons 1, 4, 5, 7)
SC.912.N.1.2 Describe and explain what characterizes science and its methods.
(Lessons 1, 4, 5, 7)
SC.912.N.1.3 Recognize that the strength of usefulness of a scientific claim is evaluated
through scientific argumentation, which depends on critical and logical thinking, and the
active consideration of alternative scientific explanations to explain the data presented.
(Lessons 1, 4, 5, 7)
SC.912.N.1.5 Describe and provide examples of how similar investigations conducted
in many parts of the world result in the same outcome.
(Lessons 1, 4, 5, 7)
SC.912.N.1.6 Describe how scientific inferences are drawn from scientific observations
and provide examples from the content being studied.
(Lessons 1, 3, 4, 5, 7)
SC.912.N.1.7 Recognize the role of creativity in constructing scientific questions,
methods and explanations.
11
(Lessons 1, 4, 5, 7)
SC.912.N.4.1 Explain how scientific knowledge and reasoning provide an empiricallybased perspective to inform society’s decision making.
(Lesson 1)
SC.912.N.4.2 Weigh the merits of alternative strategies for solving a specific societal
problem by comparing a number of different costs and benefits, such as human,
economic, and environmental.
(Lesson 1, 6)
SC.912.L.14.1 Describe the scientific theory of cells (cell theory) and relate the history of
its discovery to the process of science.
(Lesson 6)
SC.912.L.14.2 Relate structure to function for the components of plant and animal
cells.
Explain the role of cell membranes as a highly selective barrier (passive and active
transport).
(Lesson 2, 6)
SC.912.L.14.6 Explain the significance of genetic factors, environmental factors, and
pathogenic agents to health from the perspectives of both individual and public health.
(Lesson 3, 6)
SC.912.L.14.31 Describe the physiology of hormones including the different types and
the mechanisms of their action.
(Lesson 3)
SC.912.L.14.40 Describe the histology of the major arteries and veins of systemic,
pulmonary, hepatic portal, and coronary circulation.
(Lesson 2)
SC.912.L.18.2 Describe the important structural characteristics of monosaccharides,
disaccharides, and polysaccharides and explain the functions of carbohydrates
in living things.
(Lesson 2, 3)
12
Background Information
The introduction gives the rationale for conducting this unit. Regenerative medicine is
the cutting edge of medicine today, and your students can be some of the first to complete a
lab specifically designed for the high school level.
There is flexibility with the unit as well, as biology teachers are faced with the time
constraints of EOCs and AP tests. It can be done without the simulations from Science Take-out,
they just add some credibility to the scenario that someone needs a new liver. The webquest
and journal article lessons could be assigned as homework. The centerpiece, which is the liver
decellularization, can be done in two days. Time after the EOC or AP test could be utilized to
teach the unit as well.
The unit was designed to work with biology or anatomy/physiology courses, although I
will be using it with my experimental science students.
Students in grades 9-12 can complete the unit with understanding, and it probably will
work best with honors level or Advanced Placement level students. The reading of the scientific
journal article is probably the most difficult segment.
13
Bioengineered Livers in the Classroom,
You can’t Really do that?
Pre-Test
This is only a pre-test to determine how much you know before starting the unit.
Place the letter of the best choice on the line.
_____ 1.
After decellularization, what remains? A) extracellular matrix
B) cytoplasm C) cell nuclei D) cytoplasm
_____ 2.
Which chemical, made by the pancreas, is used to regulate blood glucose
levels? A) pancreatic juice B) insulin C) glycogen D) collagen
_____ 3.
Juvenile diabetes is another name for? A) Type 1 diabetes B) Type 2
diabetes C) gestational diabetes D) prediabetes
_____ 4.
Of those with diabetes, about ____% have Type 1. A) 90% B) 50%
C) 25% D) 5%
_____ 5.
Which of the following is NOT a risk factor for diabetes? A) diet
B) exercise C) education level D) genetics
_____ 6.
What is the purpose of organ decellularization? A) remove only the bad
cells B) provide a scaffold for reseeding C) provide a means of gene
therapy D) reduce organ failure in diseased individuals
_____ 7.
Which of the following is NOT a complication of diabetes? A) limb
amputation B) blindness C) increased heart disease D) increased
Alzheimer’s
_____ 8.
What is the advantage of using embryonic stem cells? A) they don’t form
teratomas B) they only become one type of cell C) they are pluripotent
D) they are not controversial
_____ 9.
Which of the following is NOT a symptom of diabetes? A) loss of
appetite B) frequent urination C) sores that heal slowly D) excessive
thirst
_____ 10.
Normal blood glucose levels are about _____ mg/dL. A) 30 B) 100
C) 200 D) 300
_____ 11.
How did researchers conclude that scaffolding was safe? A) it passed
14
OSHA guidelines B) cells grew on it C) it failed to create an immune
response D) macrophages were able to destroy it
Use the following scenario to answer questions 12-16.
Dr. Sullivan is testing an implant to determine if it can be safely placed into the
body. He has treated the implants with different types of surfactants. The implant is
surgically placed into some rats, while other rats get surgery with no implant. After 8
weeks, he looks at the levels of inflammation associated at the implant site.
_____ 12.
The amount of inflammation would be the _______. A) independent
variable B) dependent variable C) control D) constant
_____ 13.
The type of rat used would be the _____. A) independent variable
B) dependent variable C) control D) constant
_____ 14.
The type of surfactant used to treat the implant would be the _____.
A) independent variable B) dependent variable C) control D) constant
_____ 15.
The rats that received surgery but no implants would be the _____.
A) independent variable B) dependent variable C) control D) constant
_____ 16.
What is the minimum number of trials or specimens for a scientific
experiment? A) 1 B) 2 C) 3 D) 100
_____ 17.
What is the biggest disadvantage of using embryonic stem cells?
A) they can form teratomas B) they are not pluripotent C) they only
work in the original host D) there are no embryos available to get stem
cells from
_____ 18.
What happens in Type 1 diabetes? A) glucose levels drop too low
B) collagen levels are too high C) no insulin is produced D) insulin is not
used by the cells
_____ 19.
What happens in Type 2 diabetes? A) glucose levels drop too low
B) collagen levels are too high C) no insulin is produced D) insulin is not
used by the cells
_____ 20.
When would someone get gestational diabetes? A) by not exercising
B) when pregnant C) after eating a low calorie diet D) after eating a high
cholesterol diet
_____ 21.
Of those with diabetes, about ____% have Type 2. A) 90% B) 50%
15
C) 25% D) 5%
_____ 22.
Adult onset diabetes is another name for? A) Type 1 diabetes B) Type 2
diabetes C) gestational diabetes D) prediabetes
_____ 23.
How many Americans are estimated to have prediabetes? A) 1 million
B) 10 million C) 50 million D) 80 million
_____ 24.
rate
What is the best prevention for Type 1 diabetes? A) reduce your heart
by exercising less B) eat a healthier diet C) increase the amount of
fructose in the diet D) there is no prevention
_____ 25.
rate
What is the best prevention for Type 2 diabetes? A) reduce your heart
by exercising less B) eat a healthier diet C) increase the amount of
fructose in the diet D) there is no prevention
_____ 26.
How many Americans have some form of diabetes? A) 20 million
B) 30 million C) 40 million D) 50 million
_____ 27.
What is the main type of cell found in the liver? A) hepatocyte
B) macrophage C) neurofibril D) thrombocyte
_____ 28.
Which of the following is NOT a function of the liver? A) metabolizes
drugs/medicine B) stores glycogen C) digests proteins D) metabolizes
lipids
_____ 29.
Which of the following diseases is NOT associated with the liver?
A) hepatitis B) malaria C) cirrhosis D) ischemia
_____ 30.
What is the purpose of adding a detergent/surfactant to an organ during
decellularization? A) strengthen the ECM B) cause cell lysis
C) keep vascular tissue intact D) maintain viability for transplant
16
Bioengineered Livers in the Classroom,
You can’t Really do that?
Pre/Post Test Answer Key
1.
2.
3.
4.
5.
6.
7.
8.
9.
10.
11.
12.
13.
14.
15.
17
A
B
A
D
C
B
D
C
A
B
C
B
D
A
C
16.
17.
18.
19.
20.
21.
22.
23.
24.
25.
26.
27.
28.
29.
30.
C
A
C
D
B
A
B
D
D
B
A
A
C
D
B
LESSON ONE
TITLE: What is Regenerative Medicine?
KEY QUESTION(S): How is regenerative medicine different from traditional medicine? How are organs
bioengineered? What are the trends with diabetes rates in the United States?
SCIENCE CONCEPTS: Biotechnology, scientific discovery, experimental design, epidemiology.
OVERALL TIME ESTIMATE: One 50 minute class period. Extra time may be needed depending on how
quickly students are able to make their graph.
LEARNING STYLES: Visual and auditory.
VOCABULARY:
Bioengineering: Using biology, chemistry and physics to come up with real world applications for
biological problems.
Bioprinting: The use of 3D printers to produce molds using biological materials like collagen.
CDC: The U.S. Centers for Disease Control and Prevention. Part of the U.S. federal government, the CDC
tracks all things regarding public health, from viral outbreaks to obesity.
Decellularization: The process of treating organs with detergents to remove all the cells, leaving behind
the extracellular matrix (ECM).
Diabetes: Disorder in which either the body makes no insulin (Type 1) or cells aren’t able to use the
insulin properly (Type 2).
Extracellular Matrix (ECM): The non-living material that is between the cells that gives the organ its
shape, consisting mostly of collagen. Sometimes referred to as a scaffold for bioengineering organs.
Regenerative Medicine: Branch of medicine that seeks to replace defective body parts with new
bioengineered parts.
Scaffold: The non-living architectural component of an organ, consisting primarily of ECM.
Stem Cell: A cell that can differentiate/change into many different types of cells.
LESSON SUMMARY: Students will learn about advances in regenerative medicine by viewing short
video clips from researchers explaining the process of producing bioengineered organs. The CDC
database will be used to create a graph about diabetes trends in the U.S. and calculate percent increase
for several groups.
18
STUDENT LEARNING OBJECTIVES WITH NEXT GENERATION SUNSHINE STATE STANDARDS:
1. Describe how regenerative medicine is different from traditional medicine.
2. Explain the procedure for bioengineering an organ.
3. Use a database to create a graph, calculate percent increase, and analyze disease trends.
19
SC.912.N.1.1
Define a problem based on a specific body of knowledge, for example: biology,
chemistry, physics, and earth/space science, and do the following:
12. pose questions about the natural world,
13. conduct systematic observations,
14. examine books and other sources of information to see what is already
known,
15. review what is known in light of empirical evidence,
16. plan investigations,
17. use tools to gather, analyze, and interpret data (this includes the use of
measurement in metric and other systems, and also the generation and
interpretation of graphical representations of data, including data tables
and graphs),
18. pose answers, explanations, or descriptions of events,
19. generate explanations that explicate or describe natural phenomena
(inferences),
20. use appropriate evidence and reasoning to justify these explanations to
others,
21. communicate results of scientific investigations, and
22. evaluate the merits of the explanations produced by others.
SC.912.N.1.2
Describe and explain what characterizes science and its methods.
SC.912.N.1.3
Recognize that the strength of usefulness of a scientific claim is evaluated
through scientific argumentation, which depends on critical and logical thinking,
and the active consideration of alternative scientific explanations to explain the
data presented.
SC.912.N.1.5
Describe and provide examples of how similar investigations conducted in many
parts of the world result in the same outcome.
SC.912.N.1.6
Describe how scientific inferences are drawn from scientific observations and
provide examples from the content being studied.
SC.912.N.1.7
Recognize the role of creativity in constructing scientific questions, methods and
explanations.
SC.912.N.4.1
Explain how scientific knowledge and reasoning provide an empirically-based
perspective to inform society’s decision making.
SC.912.N.4.2
Weigh the merits of alternative strategies for solving a specific societal problem
by comparing a number of different costs and benefits, such as human,
economic, and environmental.
MATERIALS:
ESSENTIAL:
 1 copy of “What is Regenerative Medicine? An Introductory WebQuest” for each
student.
 Internet access. The preferred method is to have each student at their own computer,
with headphones for sound. Another option would be to use one classroom computer
with a data projector and view the clips together as a class. A third option would be to
allow students to use smart phones to access the online content.
 Calculator, although the math could be done on paper. Most student smart phones can
also double as a calculator.
SUPPLEMENTAL:
 Colored pencils for graphing.
BACKGROUND INFORMATION:
Although first mentioned in 1992, it is only in the last 4-5 years that the field of regenerative medicine
has really taken off. Traditional medicine typically seeks improve the condition of damaged organs or to
alleviate symptoms, often through therapeutic drugs. Regenerative medicine is radically different in
that it seeks to replace damaged body parts with new body parts designed in a lab. As advances in
materials technology, like 3D printers, and knowledge about cell biology increase, regenerative medicine
is closer to becoming a reality. There are several labs involved with regenerative medicine in the U.S.,
with the two most prominent ones being at Wake Forest University and the McGowan Institute at the
University of Pittsburgh.
Some researchers are looking at using synthetic materials to grow cells on to produce new organs. Only
in the last couple of years have researchers found a method of using a current organ and decellularizing
it. In the process of decellularization, detergents like Triton X 100 and SDS are used to break open the
cell membranes (lysis) and flush the cellular components out. What is left behind is the extracellular
matrix (ECM), all the non-living support material which consists of collagen. The researcher now has the
perfect architectural structure, or scaffold, in place to grow new cells. Another advantage of this
method is that the vascular tissue is left intact, making it easier to supply oxygen and nutrients to all the
20
cells along with providing a better attachment point for future transplant into a patient. Where do the
new cells come from? Stem cells have the ability to differentiate or specialize into many different types
of cells. Researchers have worked with embryonic stem cells due to their wide range of differentiation
ability. Another possibility is to use stem cells found in other parts of the body, like bone marrow or
oval cells in the liver. The advantage is that if you use the patient’s own stem cells to create the new
organ, it should not be rejected by the body after transplant. The liver is considered a prime target for
bioengineering because its main cell type, the hepatocyte, performs nearly all the functions of the liver.
Organs that have highly specialized regions and cells are more difficult to produce.
The U.S. Centers for Disease Control and Prevention is part of the federal government under the
Department of Health and Human Services. Many have heard about the CDC from high profile
investigations like H1N1, SARS or from a movie like Contagion. However, the CDC is involved in all
aspects of public health, including topics like concussions and obesity. The CDC has an extensive web
site with public information about many diseases, including a database.
ADVANCE PREPARATION:
Since much of this activity involves web based links, it would be a good idea to go through the listed
links and make sure they are still active. It would also be beneficial to review how to calculate percent
increase as students will probably have questions how to do this, even advanced students. It might be
good to have a sample problem ready to illustrate how to calculate percent increase.
PROCEDURE AND DISCUSSION QUESTIONS WITH TIME ESTIMATES:
Pass out the webquest packet to each student. The first part has students watching video clips, and the
time for each clip is listed on the webquest. This should take about 15 minutes. For the next section,
students have to read short paragraphs about bioengineering an organ from Wake Forest. There are
also some very short video clips (about 20 seconds long each) that students can view along with the
paragraphs. This section should take another 10 minutes. The next section has students create a graph,
with the data given on the webquest (taken from the CDC). Some students will have questions about
scaling the graph, which are the X and Y axes, and whether to make a line graph or bar graph. Since you
are tracking the number of people with diabetes over time, a line graph should be used (although the
CDC has a bar graph!). The graph should take about 10 minutes to complete. The final section has
students looking at data from the CDC site. Depending on the math ability of the students, it would be a
good idea to so a sample percent increase problem as an illustration or walk students through how to
solve question #18. While the teacher should circulate throughout the webquest, this is the section that
will have the most requests for assistance. This section should take 10-15 minutes depending on the
math ability of the students.
ASSESSMENT SUGGESTIONS:
For objective 1, student answers on questions #1-17 are the assessment.
For objective 2, student answers on questions #1-17 are the assessment.
21
For objective 3, student graph and answers to questions #18-26 are the assessment.
EXTENSIONS:
ACTIVITIES: None
LITERATURE: None
RESOURCES/REFERENCES:
Sullivan, D, Mirmalek-Sani, S, Deegan, B. Decellularization methods of porcine kidneys for whole organ
engineering using a high-throughput system. Biomaterials 2012; 33:7756-7764.
Wake Forest Institute for Regenerative Medicine, www.wakehealth.edu/WFIRM/ , accessed 6/25/2013.
McGowan Institute for Regenerative Medicine, www.mirm.pitt.edu, accessed 6/25/2013.
22
Name___________________
What is Regenerative Medicine?
An Introductory WebQuest
Have you ever heard of regenerative medicine? This is a new branch of medicine that is
seen by many as the future of medicine. To introduce regenerative medicine, go to the
following links and answer the questions.
Medicine’s Cutting Edge: Re-growing Organs at:
www.cbsnews.com/8301-3445_1623960219.html (4:08)
1. How long did it take for Mr. Spievak to regrow his fingertip? 4 weeks
2. Why are stem cells added to some people’s hearts? to grow new arteries
How It’s Made: Regenerative Medicine at:
science.discovery.com/tv-shows/how-its-made/videos/how-its-made-regenerativemedicine.htm (2:16)
3. What type of cells are added to the ear mold? cartilage cells
4. How is the ear mold made? computer controlled using synthetic polymer
One method of creating new organs is to first decellularize them. Check out what the
University of Minnesota is doing at:
www.youtube.com/watch?v=j9XzN0-TQZc (2:20)
5. What does decellularization mean? to remove the cells
6. How does the heart look different after decellularization? color is clear/white
In addition to hearts, other researchers are making lungs.
http://www.youtube.com/watch?v=ePox2uYcAos (3:09)
7. What is the biggest problem with lung transplants? donor lungs don’t match and get
rejected by the host
23
8. How would bioengineered lungs help prevent this problem? by using the host’s own
stem cells, the new lung would not be recognized as foreign or rejected
One of the leading universities studying regenerative medicine is Wake Forest. Their
Institute for Regenerative Medicine web site explains organ bioengineering. Go to:
www.wakehealth.edu/WFIRM/
On the left side, click on the link titled “The ABCs of Organ Engineering.”
Now click on the link “It all Starts with Cells.”
9. What is the advantage of using placental tissue stem cells? they are readily available
and don’t form tumors
Next, click “Making a Scaffold.”
10. What is a scaffold? the mold, shape, or structure to build off
The next link is “Bioprinting.”
11. What is bioprinting? using a 3D printer to make body parts
Now click on the link “Materials Selection.”
12. What three characteristics do scientists want in a scaffold material? compatible
with the body, promotes cell growth, degrades in the body once the tissue has grown
13. How do researchers test potential scaffold materials? by use of a high-throughput
robotic system
The next link is “Quality Assurance.”
14. If you want to see if cells are attaching to the scaffold, which type of microscope
would you use? scanning electron microscope
The final link under ABCs is “Testing Functionality.”
15. How are engineered organs tested? they are placed into an organ bath
Back on the left side of the web page, find the link “Our Research Projects.”
24
Click on “Military Applications.”
16. List the five areas that AFIRM focuses on to help wounded soldiers. burn repair,
wound healing without scarring, craniofacial reconstruction, limb reconstruction,
compartment syndrome
In the center of the page is the link “Growing Fingers and Limbs in the Lab.”
17. Why is it difficult to grow a finger in the lab? it has many different tissue types
Besides injury, there are many diseases that damage parts of the body. One disease you
will be learning more about is diabetes. The U.S. Centers for Disease Control and
Prevention (CDC) is part of our federal government and tracks various diseases from
ebola to obesity.
Go to http://www.cdc.gov/diabetes/surveillance/
Click on “Diabetes Data & Trends”
Use the data below, taken from the CDC, to create a graph showing the number of
Americans (in millions) diagnosed with diabetes. Be sure to give your graph a title and
label the X and Y axes.
25
Year
1980
1981
1982
1983
1984
1985
1986
1987
1988
1989
1990
2002
2003
2004
2005
2006
Number (in millions)
5.6
5.6
5.7
5.8
5.9
6.2
6.4
6.4
6.4
6.4
6.6
13.6
14.3
15.2
16.3
17.0
Year
1991
1992
1993
1994
1995
1996
1997
1998
1999
2000
2001
2007
2008
2009
2010
2011
Number (in millions)
6.9
7.5
7.6
8.1
8.0
8.8
9.4
10.5
11.1
12.0
12.9
17.8
19.0
20.2
20.8
20.9
18. Calculate the percent increase in diabetes from 1980 to 2011. 273%
19. Which year showed the highest number increase? 2007-2008 or 2008-2009
20. Which year showed the highest percent increase? 1995-1996 (10%)
On the left side, click on the link “National Surveillance Data.”
Now click on “Prevalence.”
Now click on “Percentage by Race.”
21. Which race has the highest percentage of diabetes? black
22. Which race has the highest percent increase of diabetes from 1980 to 2011? white
Go back to the “Prevalence” page and now click on “Percentage by Sex.”
23. Over the last 30 years, what trend has developed in diabetes rates between men
and women? men are getting diabetes slightly more than women
Go back to the “Prevalence” page and now click on “Percentage by Education.”
24. What is the correlation between education level and diabetes rates? the lower the
education level, the higher the diabetes rate
On the upper left side, find the link “State Surveillance Data.”
26
25. How many Floridians were diagnosed with diabetes in 2010? 1,488,000
The Florida Department of Health maintains the web site www.floridacharts.com.
Go to the site, and type in “Diabetes” in the search box. Scroll down to find the link
“Adults with Diagnosed Diabetes.”
26. What is the diabetes rate for your county? varies by county
27
Name___________________
What is Regenerative Medicine?
An Introductory WebQuest
Have you ever heard of regenerative medicine? This is a new branch of medicine that is
seen by many as the future of medicine. To introduce regenerative medicine, go to the
following links and answer the questions.
Medicine’s Cutting Edge: Re-growing Organs at:
www.cbsnews.com/8301-3445_1623960219.html (4:08)
1. How long did it take for Mr. Spievak to regrow his fingertip? ____________________
2. Why are stem cells added to some people’s hearts?
____________________________
_______________________________________________________________________
How It’s Made: Regenerative Medicine at:
science.discovery.com/tv-shows/how-its-made/videos/how-its-made-regenerativemedicine.htm (2:16)
3. What type of cells are added to the ear mold?
_________________________________
4. How is the ear mold made? _______________________________________________
One method of creating new organs is to first decellularize them. Check out what the
University of Minnesota is doing at:
www.youtube.com/watch?v=j9XzN0-TQZc (2:20)
5. What does decellularization mean? _________________________________________
6. How does the heart look different after decellularization?
_______________________
In addition to hearts, other researchers are making lungs.
http://www.youtube.com/watch?v=ePox2uYcAos (3:09)
7. What is the biggest problem with lung transplants? ____________________________
_______________________________________________________________________
8. How would decellularized lungs help prevent this problem? _____________________
_______________________________________________________________________
28
One of the leading universities studying regenerative medicine is Wake Forest. Their
Institute for Regenerative Medicine web site explains organ bioengineering. Go to:
www.wakehealth.edu/WFIRM/
On the left side, click on the link titled “The ABCs of Organ Engineering.”
Now click on the link “It all Starts with Cells.”
9. What is the advantage of using placental tissue stem cells? ______________________
________________________________________________________________________
Next, click “Making a Scaffold.”
10. What is a scaffold? _____________________________________________________
The next link is “Bioprinting.”
11. What is bioprinting? ____________________________________________________
Now click on the link “Materials Selection.”
12. What three characteristics do scientists want in a scaffold material?
______________
________________________________________________________________________
13. How do researchers test potential scaffold materials?
__________________________
________________________________________________________________________
The next link is “Quality Assurance.”
14. If you want to see if cells are attaching to the scaffold, which type of microscope
would you use? __________________________________________________________
The final link under ABCs is “Testing Functionality.”
15. How are engineered organs tested?
________________________________________
________________________________________________________________________
29
Back on the left side of the web page, find the link “Our Research Projects.”
Click on “Military Applications.”
16. List the five areas that AFIRM focuses on to help wounded soldiers. _____________
_______________________________________________________________________
In the center of the page is the link “Growing Fingers and Limbs in the Lab.”
17. Why is it difficult to grow a finger in the lab? _______________________________
Besides injury, there are many diseases that damage parts of the body. One disease you
will be learning more about is diabetes. The U.S. Centers for Disease Control and
Prevention (CDC) is part of our federal government and tracks various diseases from
ebola to obesity.
Go to http://www.cdc.gov/diabetes/surveillance/
Click on “Diabetes Data & Trends”
Use the data below, taken from the CDC, to create a graph showing the number of
Americans (in millions) diagnosed with diabetes. Be sure to give your graph a title and
label the X and Y axes.
30
Year
1980
1981
1982
1983
1984
1985
1986
1987
1988
1989
1990
2002
2003
2004
2005
2006
Number (in millions)
5.6
5.6
5.7
5.8
5.9
6.2
6.4
6.4
6.4
6.4
6.6
13.6
14.3
15.2
16.3
17.0
Year
1991
1992
1993
1994
1995
1996
1997
1998
1999
2000
2001
2007
2008
2009
2010
2011
Number (in millions)
6.9
7.5
7.6
8.1
8.0
8.8
9.4
10.5
11.1
12.0
12.9
17.8
19.0
20.2
20.8
20.9
18. Calculate the percent increase in diabetes from 1980 to 2011.
___________________
19. Which year showed the highest number increase? ____________________________
20. Which year showed the highest percent increase? ____________________________
On the left side, click on the link “National Surveillance Data.”
Now click on “Prevalence.”
Now click on “Percentage by Race.”
21. Which race has the highest percentage of diabetes?
___________________________
22. Which race has the highest percent increase of diabetes from 1980 to 2011?
_______
Go back to the “Prevalence” page and now click on “Percentage by Sex.”
23. Over the last 30 years, what trend has developed in diabetes rates between men
and women?
________________________________________________________________
Go back to the “Prevalence” page and now click on “Percentage by Education.”
31
24. What is the correlation between education level and diabetes rates?
______________
________________________________________________________________________
On the upper left side, find the link “State Surveillance Data.”
25. How many Floridians were diagnosed with diabetes in 2010? ___________________
The Florida Department of Health maintains the web site www.floridacharts.com.
Go to the site, and type in “Diabetes” in the search box. Scroll down to find the link
“Adults with Diagnosed Diabetes.”
26. What is the diabetes rate for your county?
___________________________________
32
LESSON TWO
TITLE: What is a Liver? A Beginner’s Guide
KEY QUESTION(S): Why is the liver important? What are some diseases of the liver?
SCIENCE CONCEPTS: Structure and Function of Biological Systems, Mechanism of Disease
OVERALL TIME ESTIMATE: One 50 minute class period.
LEARNING STYLES: Visual and auditory.
VOCABULARY:
Artery: Blood vessel that carries oxygen rich blood away from the heart.
Bile: Fluid produced by the liver and stored in the gall bladder, breaks down fats.
Cirrhosis: After chronic liver disease, fibrous tissue replaces liver tissue and liver loses function.
Fibrosis: Fibrous tissue replaces liver tissue, leads to cirrhosis.
Hepatitis: General term for inflammation of the liver, can be caused by several pathogens Hepatocyte:
Main type of cell found in the liver. Not highly specialized, can perform most liver functions.
Kuppfer cell: A macrophage of the liver, it can clear out debris, old red blood cells and pathogens.
Lobule: Small functional unit of the liver where blood flows into the sinusoids or spaces between the
blood vessels.
Regenerate: The ability to regrow and function. The liver is the only internal organ that can regrow.
Vein: Blood vessel that carries oxygen poor blood towards the heart.
LESSON SUMMARY: This lesson will introduce students to the basic anatomy of the liver, its major
functions and diseases that affect the liver. A Powerpoint presentation with accompanying student note
packet will be the main method of delivery.
STUDENT LEARNING OBJECTIVES WITH NEXT GENERATION SUNSHINE STATE STANDARDS:
1. Describe the major anatomical features of the liver.
2. Describe the functions of the liver.
3. Explain the conditions of hepatitis, fibrosis, cirrhosis and cancer as disease progression in the
liver.
SC.912.L.14.2
33
Relate structure to function for the components of plant and animal cells.
Explain the role of cell membranes as a highly selective barrier (passive and
active transport).
SC.912.L.14.40 Describe the histology of the major arteries and veins of systemic, pulmonary,
hepatic portal, and coronary circulation.
SC.912.L.18.2
Describe the important structural characteristics of monosaccharides,
disaccharides, and polysaccharides and explain the functions of carbohydrates
in living things.
MATERIALS:
ESSENTIAL:
 Classroom computer with data projector.
 Copies of student note companion.
SUPPLEMENTAL:
 Liver diagram.
 Human torso model with liver.
 Fresh beef liver from the grocer.
BACKGROUND INFORMATION:
The liver is the largest internal organ in the body, weighing 3-4 pounds, and is second only to the skin in
overall size. It is divided into four sections or lobes, but each section is not specialized. In fact, one of
the reasons why the liver is a good candidate for bioengineering is that has one main type of cell, called
a hepatocyte. Hepatocytes make up 80% of the liver by volume, and can carry out all liver functions.
Livers have the unique ability to regenerate, or grow back. If up to 80% of the liver is surgically
removed, it will grow back to its original size with 30 days. Specialized Kuppfer cells are macrophages
that clear away debris, old red blood cells and pathogens. The gall bladder is attached to the liver and
stores bile. One fourth of the cardiac output enters the liver through the hepatic artery. Blood returns
to the heart by the hepatic vein. The portal vein enters the underside, bringing in nutrient rich blood
from the intestines. A liver lobule is the functional unit of the liver, where all the action takes place.
Blood moves from vessels into the spaces or sinusoids, and is acted upon by the hepatocytes.
The liver has several functions, and you can’t survive without a liver. Bile is produced by the liver, and is
used to break down lipids/fats. Blood sugar levels are assisted by the liver, as it takes glucose and
converts it to glycogen for storage. Blood clotting proteins are also produced by the liver. Lipids are
metabolized by the liver, along with any type of drug or medication. In fact, before receiving strong
medicines, doctors usually perform a liver function test to determine if the liver can handle the
34
additional stress. Another important function is the detoxification of harmful substances like alcohol,
converting them into waste products that can be excreted by the body.
There are several diseases that affect the liver. Hepatitis is the general term for inflammation of the
liver, and there can be several causes. Many people think about viral hepatitis, with hepatitis A, B and C.
Hepatitis A usually enters the body from contaminated food. Symptoms occur between two and six
weeks after infection, and it usually lasts for four weeks. Hepatitis B is transmitted by blood or body
fluids, and therefore can be sexually transmitted. Hepatitis B is very common in parts of Asia and Africa,
with about 350 million chronic sufferers. Hepatitis C is also blood borne, and is most commonly
contracted by intravenous drug usage or poorly sanitized medical equipment. Most people respond to
drug treatments for Hepatitis C, but 80% of those exposed will develop a chronic condition if not
treated. All varieties of viral hepatitis harm the liver by having the virus replicate in the hepatocytes. In
addition to viruses, other causes of hepatitis include drugs, alcohol and even parasites like worms.
When the hepatitis goes into a chronic state, the body responds by making fibrous tissue that replaces
liver tissue. The liver is now scarred and doesn’t function as well. Further deterioration results in
cirrhosis, with a great deal of fibrous tissue. At this point, patients need a liver transplant. Many times
cancer sets in after cirrhosis.
ADVANCE PREPARATION:
Prior to this lesson, the instructor should be very familiar with the anatomy of the liver. Copies of the
student notes should be made, along with a liver diagram. There are several sites that have liver
diagrams available, some are free. Many schools have a human torso model with removable organs.
This would be an excellent way to show students the location and relative size of the liver. Another
possibility is to get a fresh beef liver from a grocery store.
PROCEDURE AND DISCUSSION QUESTIONS WITH TIME ESTIMATES:
The opening activity should take 5 minutes. If you have a human torso model available, ask for a
volunteer to locate the liver. List the following organs: stomach, liver, small intestine, heart, lungs. Ask
students to rank them in order of critical need to the body. The liver is the only organ for which there is
no artificial device to keep a person alive! Ask students to write down what the liver does for them.
After 2-3 minutes, discuss student answers. Depending on your students, there are some fairly graphic
pictures online of people with cirrhosis. This could be an attention grabber, and you could ask students
why the person has that appearance.
Pass out the student note forms. Go through the PowerPoint presentation on the liver. This will take
about 30 minutes.
Pass out the liver diagram and have students color code it. This will take about 10 minutes.
Ask students to answer the following in their notebook: “ What are the two main types of cells found in
the liver?”; “List three major functions of the liver.”; and ”How does cirrhosis develop?” Circulate as
students write down their answers and check for understanding.
35
ASSESSMENT SUGGESTIONS:
For objective 1, student answer to notebook question #1 is the assessment.
For objective 2, student answer to notebook question #2 is the assessment.
For objective 3, student answer to notebook question #3 is the assessment.
EXTENSIONS:
ACTIVITIES: None
LITERATURE: None
RESOURCES/REFERENCES:
Thibadeau, G. Anthony’s Textbook of Anatomy & Physiology. Mosby Publishing, 1996.
Brown University,
www.biomed.brown.edu/Courses/B1108/B1108_2002_Groups/liver/webpage/Normal Liver.htm ,
accessed 6/25/2013
36
What is a Liver?
I. Liver
A. Largest
1. weighs
2. four
3. gall bladder
4. you can’t survive
5. can
a. 80%
b. grows back
B. Cell
1.
a. make up 80%
b. perform most
c. aren’t
2. Kupffer
a.
b. make up 15%
C. Circulation
1. hepatic
a. 25% of blood
b. oxygen
2. portal
a. brings in blood
b. low
c. nutrient
D.
1. functional unit
2. many small
3. great surface
37
II. Liver
A. Produces
1. breaks down
2. stored in the
B. Converts glucose to glycogen,
C. Metabolizes
D.
1. drugs/
2.
E. Produces blood
F. Removes old
III. Liver
A. Hepatitis – general term
1.
a. hepatitis A –
b. hepatitis B –
c. hepatitis C –
2. drugs/
3.
B.
1. hepatitis leads to
2. scarring of the liver,
C.
1. condition after chronic
2. little
3. liver
D. Cancer
38
LESSON THREE
TITLE: Diagnosing Diabetes
KEY QUESTION(S): What is diabetes? What are the different types of diabetes?
SCIENCE CONCEPTS: Epidemiology, homeostasis, biochemistry.
OVERALL TIME ESTIMATE: One 50 minute class period.
LEARNING STYLES: Visual, auditory and kinesthetic.
VOCABULARY:
Diabetes: Disorder in which either the body makes no insulin (Type 1) or cells aren’t able to use the
insulin properly (Type 2).
Glucose: A monosaccharide or simple sugar (C6H12O6), it is the major fuel for cellular respiration.
Homeostasis: Maintaining a balance, or regulating the environment in the cells or body as a whole.
Hormone: A chemical released by a cell, gland or organ and affects another part of the body.
Insulin: A hormone made in the pancreas that allows cells to absorb glucose.
Pancreas: An endocrine/digestive gland that produces pancreatic juice and insulin.
Receptor: A molecule on the cell membrane that receives chemical signals that direct the cell.
LESSON SUMMARY: Students will learn about diabetes by completing an information sheet. They will
also conduct a simulated glucose tolerance test to determine what type (if any) of diabetes their patient
has.
STUDENT LEARNING OBJECTIVES WITH NEXT GENERATION SUNSHINE STATE STANDARDS:
1. Compare Type 1 and Type 2 diabetes.
2. Explain how the glucose tolerance test can detect diabetes.
SC.912.L.14.6
Explain the significance of genetic factors, environmental factors, and
pathogenic agents to health from the perspectives of both individual and public
health.
SC.912.L.14.31 Describe the physiology of hormones including the different types and the
mechanisms of their action.
39
SC.912.N.1.6
Describe how scientific inferences are drawn from scientific observations and
provide examples from the content being studied.
MATERIALS:
ESSENTIAL:
 One lab packet per group of STO-117 Diagnosing Diabetes from science take-out. All
supplies are included in the packet.
 One copy of the student worksheet in the packet, per student.
SUPPLEMENTAL:
BACKGROUND INFORMATION:
As seen in the previous day’s assignment, the incidence of diabetes has increased tremendously over
the last 30 years. Genetics does play a role, but for many people a diet high in fats/lipids and lack of
exercise lead to Type 2 diabetes.
Dr. Seh-Hoon Oh, who works in the Petersen Lab, is currently experimenting with what he believes is a
factor in Type 1 diabetes. He has found a new protein, named Islet Homeostasis Protein (IHoP), which
appears to be necessary for blood glucose level regulation.
ADVANCE PREPARATION:
Since this simulation comes pre-packaged, the teacher should go through a trial run before the lab. If
you plan to reuse the materials, the graphics should be laminated and cut out. Have students place the
animations on their worksheet without gluing them down. The teacher will have to visually inspect the
worksheet for accuracy during the simulation.
PROCEDURE AND DISCUSSION QUESTIONS WITH TIME ESTIMATES:
See attached PDF from Science take-out.
ASSESSMENT SUGGESTIONS:
For objective 1, student graphic organizer is the assessment.
For objective 2, student answers on questions #5-10 are the assessment.
EXTENSIONS:
ACTIVITIES:

40
Have students take a diabetes risk test from the American Diabetes Association. It takes
one minute and they can use their smart phone to access it at:
www.diabetes.org/diabetes-basics/prevention/diabetes-risk-test
LITERATURE: None
RESOURCES/REFERENCES:
Oh, S, Darwiche, H. Characterization of a novel functional protein in the pancreatic islet: IHoP regulation
of glucagon synthesis in alpha-cells. Pancreas 2012 41(1), 22-30.
The American Diabetes Organization, www.diabetes.org , accessed 6/20/2013.
41
LESSON FOUR
TITLE: Reading a Scientific Journal
KEY QUESTION(S): How do scientists conduct a high level experiment? How do scientists describe their
experiment?
SCIENCE CONCEPTS: Experimental design.
OVERALL TIME ESTIMATE: One 50 minute class period.
LEARNING STYLES: Visual
VOCABULARY:
Allograft gold standard: An organ transplant between members of the same species, like from one
person to another.
Anesthetized: To be placed under anesthesia, or to lose sensation.; performed before surgery so the
patient won’t feel pain.
Apoptotic: When cells die off.
Cannulated: Placing a small tube (cannula) into the body.
Cytokine: Small protein molecules that are associated with inflammation.
Euthanized: Intentionally killing an organism.
Heparinized: Treated with an anticoagulant (heparin) to prevent blood clotting.
Hepatic: Refers to the liver.
Histologically: Refers to the study of tissues.
Immunogenicity: The ability of a substance to cause an immune response.
Lysis: Breaking apart a cell.
Macrophage: Large cells that perform phagocytosis, or ‘eating’ of foreign particles.
Perfused: To permeate an organ with a fluid.
Porcine: Refers to a pig.
Proinflammatory: Promotes inflammation.
Protocol: A set of procedures or steps to be followed.
42
Therapeutic: Has the ability to heal, reduce symptoms.
Sham Operated: A fake or false operation. Used as a control in the context of this paper.
Tonal: Refers to different shade of color in the context of this paper.
Xenogenic: From different species, as in a transplant from a pig into a rat.
LESSON SUMMARY: Students will be reading a scientific journal article that relates to the liver
decellularization experiment they will be performing the next day.
STUDENT LEARNING OBJECTIVES WITH NEXT GENERATION SUNSHINE STATE STANDARDS:
1. Describe how scientists set up a high level experiment.
2. Examine data to determine if it supports the hypothesis.
43
SC.912.N.1.1
Define a problem based on a specific body of knowledge, for example: biology,
chemistry, physics, and earth/space science, and do the following:
23. pose questions about the natural world,
24. conduct systematic observations,
25. examine books and other sources of information to see what is already
known,
26. review what is known in light of empirical evidence,
27. plan investigations,
28. use tools to gather, analyze, and interpret data (this includes the use of
measurement in metric and other systems, and also the generation and
interpretation of graphical representations of data, including data tables
and graphs),
29. pose answers, explanations, or descriptions of events,
30. generate explanations that explicate or describe natural phenomena
(inferences),
31. use appropriate evidence and reasoning to justify these explanations to
others,
32. communicate results of scientific investigations, and
33. evaluate the merits of the explanations produced by others.
SC.912.N.1.2
Describe and explain what characterizes science and its methods.
SC.912.N.1.3
Recognize that the strength of usefulness of a scientific claim is evaluated
through scientific argumentation, which depends on critical and logical thinking,
and the active consideration of alternative scientific explanations to explain the
data presented.
SC.912.N.1.5
Describe and provide examples of how similar investigations conducted in many
parts of the world result in the same outcome.
SC.912.N.1.6
Describe how scientific inferences are drawn from scientific observations and
provide
examples from the content being studied.
SC.912.N.1.7
Recognize the role of creativity in constructing scientific questions, methods and
explanations.
MATERIALS:
ESSENTIAL:
 One copy of the scientific journal worksheet for each student.
 One copy of the scientific journal for each student.
SUPPLEMENTAL:
 Highlighters to mark main ideas or areas that need clarification.
BACKGROUND INFORMATION:
Reading a scientific journal article can be very frustrating. The articles are written to an audience highly
versed in the jargon of their specialty, so even someone with a good science background could have
difficulty. At a previous internship, the first thing I had to do was to read an article about LRH-1. I
literally sat in front of a computer with Wikipedia open just to try and make sense of it. It took me
several times reading the article before I had a grasp about what the experiment entailed. This activity
will challenge the high level students, but if they plan to advance in science, reading articles is a must.
The main idea for this article was to explain an experiment done with ECM matrix. The researchers
hypothesized that if properly treated, it wouldn’t make a difference if the ECM scaffold came from the
same species for transplant. To gauge if the scaffold is being rejected, researchers look for an immune
response by things like increased levels of white blood cells, lymphocytes and monocytes at the scaffold
site. Previous experiments with cross species (xenogeneic) transplants did show an immune response,
but the scaffold had cells seeded on it. This experiment placed only the scaffold, without any cells, to
determine if it would create an immune response. Implications of the experiment would determine if
scaffold material could be taken from animals like pigs, and then used to bioengineer a human organ.
The article briefly describes the organ decellularization process. The ECM scaffold was made from pig
livers, and were placed into rats. A control group of rats had ‘surgery’ but no scaffold was implanted.
This was to see if the surgery itself could cause an immune response. Pictures on page 5 of the article
do show a rat post-surgery, so be aware if you have students who might be upset with seeing that. An
immune response should take place with 28 days, so specimens were checked at 2, 7, 14 and 28 days.
44
Results showed that now immune response occurred and the researchers are now confident that they
can proceed with pig livers to provide the ECM scaffold to bioengineer a liver.
ADVANCE PREPARATION:
The student pre-reading vocabulary worksheet was given out two days prior to the reading of the
article. The teacher should read the article carefully a couple of times so that they are confident
answering the questions from the students. In addition, the teacher should review the journal article
worksheet answer key to see where the answers are in text.
PROCEDURE AND DISCUSSION QUESTIONS WITH TIME ESTIMATES:
The first thing to do is to collect the pre-reading vocabulary sheet previously assigned.
There are really two ways of completing this assignment. If you have advanced readers, the article can
be assigned and given the class period to complete.
Another possibility is to have students read one paragraph or section at a time, and then ask for
volunteers to summarize the content in their own words. So many ‘reading strategies’ I see students
use are for finding the answer, not understanding what was read. While this method of group reading
may take longer, more students will probably understand the article more and be in a better position for
the experiment the next day.
ASSESSMENT SUGGESTIONS:
For objective 1, questions #1-4 and #18 are the assessment.
For objective 2, questions #5, #20 and #22 are the assessment.
EXTENSIONS:
ACTIVITIES: None
LITERATURE: Several articles are listed in the bibliography.
RESOURCES/REFERENCES:
Mirmalek-Sani, S, Sullivan, D, Zimmerman, C. Immunogenicity of decellularized porcine liver for
bioengineered hepatic tissue. Pathology, August 2013.
Name_____________________
Scientific Journal Article
Vocabulary List
45
The following words are important to know in order to understand the scientific
journal article to be read in class. Use any source to define the words, but it is best to
put the definition into your own words.
1. allograft gold standard An organ transplant between members of the same species,
like from one person to another.
2. anesthetized To be placed under anesthesia, or to lose sensation. Performed before
surgery so the patient won’t feel pain.
3. apoptotic When cells die off.
4. cannulated Placing a small tube (cannula) into the body.
5. cytokine Small protein molecules that are associated with inflammation.
6. euthanized Intentionally killing an organism.
7. heparinized Treated with an anticoagulant (heparin) to prevent blood clotting.
8. hepatic Refers to the liver.
9. histologically Refers to the study of tissues.
10. immunogenicity The ability of a substance to cause an immune response.
46
11. lysis Breaking apart a cell.
12. macrophage Large cells that perform phagocytosis, or ‘eating’ of foreign particles.
13. perfused To permeate an organ with a fluid.
14. porcine Refers to a pig.
15. proinflammatory Promotes inflammation.
16. protocol A set of procedures or steps to be followed.
17. therapeutic Has the ability to heal, reduce symptoms.
18. sham operated A fake or false operation. Used as a control in the context of this paper.
19. tonal Refers to different shade of color in the context of this paper.
20. xenogenic From different species, as in a transplant from a pig into a rat.
Name___________________
Scientific Journals
47
Reading a scientific journal article can be difficult. The articles are written to
explain exactly why the scientist did an experiment, how it was done, data collected and
what the data means. This is called technical writing, and is a career that is currently in
high demand. Many scientists aren’t great writers, and there are also many scientists
who speak a first language other than English. A goal for most scientists is to get their
research published, which helps them secure their job and assists in getting future
grants. The article you are reading, “Immunogenicity of Decellularized Porcine Liver for
Bioengineered Hepatic Tissue” was written by David Sullivan, who currently works in the
Petersen lab at the University of Florida. At the time of writing this, David was a student
only a few years older than most high school students. The Petersen lab has five PhD
scientists along with a few graduate students assisting on the various projects. In a
typical year, they will get 3-4 papers published.
The first part of the paper is the abstract, which summarizes the experiment.
You may want to have Wikipedia open to find out the meaning of technical terms you
are not familiar with. Answer the following in your own words, do not just copy text!
1. What was the purpose of this study?
To see if larger pig livers could be used instead of rat livers, and then to see if
the scaffold implant caused an immune response.
2. What is the hypothesis? (not listed, but can be inferred)
If scaffold from pigs are placed into a rat, then no immune response will be
produced.
3. Which two chemicals were used to alter the liver?
Triton X 100 and SDS
4. What was done with the pig liver scaffold?
The scaffolds were placed into rats. (They also grew cells on them.)
5. Did the researchers get results that supported their hypothesis? yes
The next section of the paper reviews literature, or other studies that relate to
their experiment.
6. How many sources did the authors use to write their paper? 51
7. Why are scientists attempting to bioengineer livers?
There is a shortage of human livers available for transplant.
8. Why are cytotoxic chemicals and DNA removed from the scaffold?
These can trigger an immune response.
48
9. What organs have successfully been decellularized?
heart, lungs, bladder, intestines, kidney, liver
10. Why was the scaffold injected into the host without any cells attached?
To determine if the scaffold itself would trigger an immune response.
(Remember, they are taking scaffold from a pig and putting it into a rat.)
The materials and methods section of the paper list all of the materials used,
where they were obtained from, and a summary of the procedures. There is usually a
separate “Supplemental Materials” attachment that details the methods step by step.
Remember, another scientist should be able to follow the procedure and replicate the
results.
11. How large were the pigs that ‘donated’ their livers? 20-25 kg
12. What advantage is there to using Masson’s trichrome stain instead of H&E staining?
Has three colors, with collagen showing up as blue.
13. What were the liver scaffolds placed into? male Fischer F344 rats
14. How long were the scaffolds left in? up to 28 days
The results section presents the data obtained from the experiment. Data
tables, graphs, and pictures all contain important information.
15. Compare how the livers look before and after decellularization. (FIG 1)
The original liver was dark red, while the decellularized liver was clear.
(It helps to have a color copy of the article, if not you might be able to show
pics.)
16. After decellularization, what is the main component of the scaffold?
collagen
17. Did rat cells start to live on the implanted scaffold? yes
18. Why were some rats operated on, but no scaffold implanted?
This was a control group. Researchers wanted to see if an immune response
would be triggered just from the surgery itself.
19. What happened to the white blood cell count over 28 days?
The white blood cell levels dropped.
49
20. How does the data support the researchers’ claim of no immune response to the
scaffold?
By looking at graphs I, J and K, all three indicators showed a drop after 28 days.
If there was an immune response occurring, all three should have increased.
The last section is the discussion, where researchers explain what the data
means. This is important because an argument must be made as to whether the data
supported the hypothesis. Based on the experimental results, direction for future
studies is also considered.
21. Why do researchers want to keep the blood vessels intact after decellularization?
Once the cells start to grow, there needs to be a way to get oxygen and nutrients
to all parts of the organ. Otherwise, there would be pockets of dead cells in the
middle. It is easier and more efficient to use the vascular structures already in
place instead of trying to create a new system. (This is a huge advantage over
using synthetic scaffolds.)
22. Did the results of the experiment support the hypothesis? Explain how/why.
The results of the experiment did support the hypothesis. The scaffold from the
pig was inserted into a rat, and new cells grew on it. There was no immune
response as indicated by the levels of white blood cells, lymphocytes and
monocytes decreasing over a 28 day period.
Bonus: Who paid for this study? National Institute of Health
50
Name_____________________
Scientific Journal Article
Vocabulary List
The following words are important to know in order to understand the scientific
journal article to be read in class. Use any source to define the words, but it is best to
put the definition into your own words.
1. allograft gold standard __________________________________________________
________________________________________________________________________
________________________________________________________________________
2. anesthetized
___________________________________________________________
________________________________________________________________________
________________________________________________________________________
3. apoptotic ______________________________________________________________
________________________________________________________________________
________________________________________________________________________
4. cannulated ____________________________________________________________
________________________________________________________________________
________________________________________________________________________
5. cytokine ______________________________________________________________
________________________________________________________________________
________________________________________________________________________
6. euthanized ____________________________________________________________
________________________________________________________________________
________________________________________________________________________
7. heparinized ____________________________________________________________
________________________________________________________________________
________________________________________________________________________
8. hepatic _______________________________________________________________
________________________________________________________________________
________________________________________________________________________
9. histologically __________________________________________________________
51
________________________________________________________________________
________________________________________________________________________
10. immunogenicity _______________________________________________________
________________________________________________________________________
________________________________________________________________________
11. lysis ________________________________________________________________
________________________________________________________________________
________________________________________________________________________
12. macrophage __________________________________________________________
________________________________________________________________________
________________________________________________________________________
13. perfused _____________________________________________________________
________________________________________________________________________
________________________________________________________________________
14. porcine ______________________________________________________________
________________________________________________________________________
________________________________________________________________________
15. proinflammatory ______________________________________________________
________________________________________________________________________
________________________________________________________________________
16. protocol _____________________________________________________________
________________________________________________________________________
________________________________________________________________________
17. therapeutic ___________________________________________________________
________________________________________________________________________
________________________________________________________________________
18. sham operated ________________________________________________________
________________________________________________________________________
________________________________________________________________________
19. tonal ________________________________________________________________
________________________________________________________________________
________________________________________________________________________
52
20. xenogenic ____________________________________________________________
________________________________________________________________________
________________________________________________________________________
53
Name___________________
Scientific Journals
Reading a scientific journal article can be difficult. The articles are written to
explain exactly why the scientist did an experiment, how it was done, data collected and
what the data means. This is called technical writing, and is a career that is currently in
high demand. Many scientists aren’t great writers, and there are also many scientists
who speak a first language other than English. A goal for most scientists is to get their
research published, which helps them secure their job and assists in getting future
grants. The article you are reading, “Immunogenicity of Decellularized Porcine Liver for
Bioengineered Hepatic Tissue” was written by David Sullivan, who currently works in the
Petersen lab at the University of Florida. At the time of writing this, David was a student
only a few years older than most high school students. The Petersen lab has five PhD
scientists along with a few graduate students assisting on the various projects. In a
typical year, they will get 3-4 papers published.
The first part of the paper is the abstract, which summarizes the experiment.
You may want to have Wikipedia open to find out the meaning of technical terms you
are not familiar with. Answer the following in your own words, do not just copy text!
1. What was the purpose of this study?
________________________________________
________________________________________________________________________
________________________________________________________________________
2. What is the hypothesis? (not listed, but can be inferred)
________________________
________________________________________________________________________
________________________________________________________________________
3. Which two chemicals were used to alter the liver? _____________________________
________________________________________________________________________
4. What was done with the pig liver scaffold? ___________________________________
________________________________________________________________________
________________________________________________________________________
5. Did the researchers get results that supported their hypothesis?
___________________
The next section of the paper reviews literature, or other studies that relate to
their experiment.
54
6. How many sources did the authors use to write their paper?
_____________________
7. Why are scientists attempting to bioengineer livers? ___________________________
________________________________________________________________________
________________________________________________________________________
8. Why are cytotoxic chemicals and DNA removed from the scaffold? _______________
________________________________________________________________________
________________________________________________________________________
9. What organs have successfully been decellularized? ___________________________
________________________________________________________________________
10. Why was the scaffold injected into the host without any cells attached?
___________
________________________________________________________________________
________________________________________________________________________
The materials and methods section of the paper list all of the materials used,
where they were obtained from, and a summary of the procedures. There is usually a
separate “Supplemental Materials” attachment that details the methods step by step.
Remember, another scientist should be able to follow the procedure and replicate the
results.
11. How large were the pigs that ‘donated’ their livers?
___________________________
12. What advantage is there to using Masson’s trichrome stain instead of H&E staining?
________________________________________________________________________
________________________________________________________________________
13. What were the liver scaffolds placed into? __________________________________
14. How long were the scaffolds left in? _______________________________________
The results section presents the data obtained from the experiment. Data
tables, graphs, and pictures all contain important information.
15. Compare how the livers look before and after decellularization. (FIG 1) ___________
________________________________________________________________________
________________________________________________________________________
55
16. After decellularization, what is the main component of the scaffold? _____________
________________________________________________________________________
17. Did rat cells start to live on the implanted scaffold? ___________________________
18. Why were some rats operated on, but no scaffold implanted? ________________
________________________________________________________________________
________________________________________________________________________
19. What happened to the white blood cell count over 28 days?
_____________________
________________________________________________________________________
20. How does the data support the researchers’ claim of no immune response to the
scaffold? ________________________________________________________________
________________________________________________________________________
________________________________________________________________________
The last section is the discussion, where researchers explain what the data
means. This is important because an argument must be made as to whether the data
supported the hypothesis. Based on the experimental results, direction for future
studies is also considered.
21. Why do researchers want to keep the blood vessels intact after decellularization?
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
22. Did the results of the experiment support the hypothesis? Explain how/why.
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
________________________________________________________________________
Bonus: Who paid for this study? _____________________________________________
56
LESSON FIVE
TITLE: Bioengineer Me a Liver!: Part One
KEY QUESTION(S): How do you decellularize a liver?
SCIENCE CONCEPTS: Experimental design
OVERALL TIME ESTIMATE: One 50 minute class period for part one, with an additional 50 minute class
period for part two (Lesson Seven).
LEARNING STYLES: Visual, auditory, and kinesthetic.
VOCABULARY:
Constants: Factors that are kept the same throughout the experiment.
Control: Group that is not treated with the independent variable. Used to measure results against.
Dependent Variable: The change you are looking for, what you are measuring.
Experimental Group: Group that is treated with the independent variable.
Hypothesis: Prediction for the results of the experiment, best written as an “if-then” statement and
based on something measureable.
SDS: Sodium dodecyl sulfate, a detergent used to cause cell lysis.
Triton X 100: A non-ionic detergent used to cause cell lysis.
LESSON SUMMARY: Students will set up a controlled experiment to decellularize a chicken liver.
STUDENT LEARNING OBJECTIVES WITH NEXT GENERATION SUNSHINE STATE STANDARDS:
1. List the steps of the scientific method.
2. Describe how to set up a controlled experiment.
SC.912.N.1.1
57
Define a problem based on a specific body of knowledge, for example: biology,
chemistry, physics, and earth/space science, and do the following:
34. pose questions about the natural world,
35. conduct systematic observations,
36. examine books and other sources of information to see what is already
known,
37. review what is known in light of empirical evidence,
38. plan investigations,
39. use tools to gather, analyze, and interpret data (this includes the use of
measurement in metric and other systems, and also the generation and
interpretation of graphical representations of data, including data tables
and graphs),
40. pose answers, explanations, or descriptions of events,
41. generate explanations that explicate or describe natural phenomena
(inferences),
42. use appropriate evidence and reasoning to justify these explanations to
others,
43. communicate results of scientific investigations, and
44. evaluate the merits of the explanations produced by others.
SC.912.N.1.2
Describe and explain what characterizes science and its methods.
SC.912.N.1.3
Recognize that the strength of usefulness of a scientific claim is evaluated
through scientific argumentation, which depends on critical and logical thinking,
and the active consideration of alternative scientific explanations to explain the
data presented.
SC.912.N.1.5
Describe and provide examples of how similar investigations conducted in many
parts of the world result in the same outcome.
SC.912.N.1.6
Describe how scientific inferences are drawn from scientific observations and
provide examples from the content being studied.
SC.912.N.1.7
Recognize the role of creativity in constructing scientific questions, methods and
explanations.
MATERIALS:
ESSENTIAL:
 Raw chicken livers. One chicken liver could be used for 5-6 groups, but if you want to
see the vascular tissue it is best to have one chicken liver for each lab group. Most
grocers carry chicken livers, about $3 for 10-15 chicken livers
 Lab packet, one per student.
 Camera or smart phone, one per group.
 Flask, 1 L preferred. At least one per class.
 Stir plate with stir bar. One per flask.
 Scalpel or scissors, one per group.
 Dissection tray, one per group.
58




Ruler, one per group.
Colored pencils.
Triton X 100 1% solution. Can be purchased from several science supply companies. If
using one 1 L flask, you will need 1 L of Triton X 100 1% solution.
SDS .1% solution. Can be purchased from several science supply companies. If using
one 1 L flask, you will need 1 L of SDS .1% solution.
SUPPLEMENTAL:
BACKGROUND INFORMATION:
After seeing video clips and reading about whole organ decellularization, this is the day students will
undertake the process. It is assumed that students have already been taught the scientific method
(Don’t they get that every year since 3rd grade?) but the beginning of the lab serves as a good review. In
the Advanced Placement science courses, one of the Free Response Questions typically asked students
to describe setting up a controlled experiment. Researchers use a peristaltic pump to deliver the Triton
X and SDS solutions through the blood vessels of the organs. Your students will need to use smaller
pieces of liver to expose it to the detergents.
I did try the experiment with just SDS (the Triton X 100 is a little bit expensive) and let it stir in solution
for 3 days. The liver sections did turn color, but not nearly as dramatic as with the Triton X 100. I also
tried to use a 10% baby shampoo solution (similar to the cheap DNA extraction labs) but it didn’t work
very well. There are other types of detergents that may work, and that could be another independent
variable for students to work with. If you have access to several stir plates, you might consider making
the experiment better by offering different choices of detergents (or concentrations as Triton X 100
could be made in 1%, 3% and 5% solutions )as independent variables.
Pictures of results of my experiment are posted at the end of this lesson.
ADVANCE PREPARATION:
The biggest problem for advanced preparation is locating a stir plate/stir bar. Since you aren’t using a
peristaltic pump, the constant stirring action helps get the detergents into the liver sections. Triton X
100 is extremely viscous and will take few hours to go into solution, so don’t plan on mixing it right
before the lab. The Triton X 100 will sink to the bottom of the flask and requires vigorous stirring to go
into solution. Once in solution it is fine. It is best to order the SDS already in solution. If you order it in
the powder form, it should be mixed under a fume hood since it goes airborne and up your nose (an
irritant). The SDS should also be mixed a day before the lab. Raw chicken livers need to be refrigerated
and do have a slight odor. You might cover the flask with aluminum foil to reduce odors coming out
while decellularizing.
59
PROCEDURE AND DISCUSSION QUESTIONS WITH TIME ESTIMATES:
It will be important to use time wisely, especially at the beginning of the lab period. Students should get
into their lab groups and get a dissection tray, ruler, scalpel and raw chicken liver. Have students use a
camera or smart phone to take pictures of the liver from several angles/sides. Each group then needs to
cut out a section of liver about 1 cm x 1 cm. Larger sections may not get fully decellularized. It looks
better if you can see the vascular tissue, so the best place to get your section is from the top/bottom
center of the liver. This should be done in the first five minutes of class, because you need to change
solutions after 40 minutes. Place the liver sections into the 1 L flask with about 500 mL of Triton X 100
1% solution. Students probably will not get to identify the ‘their’ liver section after decellularization.
The solution will start to turn brown from the blood. Insert the stir bar and place on the stir plate for 40
minutes. After 40 minutes, pour the solution out of the flask (keep the liver sections!) and put in 500 mL
of fresh Triton X 100 1% solution. The time doesn’t have to be exact, so if the solution gets changed at
30 minutes or 60 minutes, you will still get results.
While the liver sections are stirring in Triton X, students can answer questions #1-6 on the lab packet.
Emphasize that the questions will be answered AFTER the liver sections get cut and placed into solution.
The teacher should circulate to answer questions students might have. I find that students often
confuse constants with controls.
ASSESSMENT SUGGESTIONS:
For objective 1, student answers to question #1 is the assessment.
For objective 2, students’ answers to questions #2-5 are the assessment.
EXTENSIONS:
ACTIVITIES: None
LITERATURE: None
RESOURCES/REFERENCES:
Sullivan, D, Mirmalek-Sani, S, Deegan, B. Decellularization methods of porcine kidneys for whole organ
engineering using a high-throughput system. Biomaterials 2012; 33:7756-7764.
60
Untreated chicken liver sections
Liver starting to turn clear in Triton X 100
61
Decellularized chicken liver sections after 24 hours
Note the blood vessels still intact
62
Name___________________
Bioengineering a Liver
Now that you have learned about regenerative medicine, the role of the liver
and diseases of the liver, it is time to take the first step in making a new liver. This is
called decellularization, which strips away all the cells leaving only behind the
extracellular matrix which is then used as a scaffold to make the new liver.
Materials:
fresh chicken liver
flask
stir plate
SDS solution
camera
ruler
scalpel
stir bar
Triton X solution
colored pencils
dissection tray
Procedure:
1.
2.
3.
4.
5.
6.
7.
8.
Get a fresh chicken liver from your teacher.
Take several pictures of the liver.
Use the scalpel to cut a piece of liver no larger than 1 cm x 1 cm. For best
results, use a section where there are veins and arteries.
Choose which flask to put your liver piece into.
Complete part one of the packet while you wait.
After 40 minutes, empty the solution and put 500 mL of new solution into the
flask.
Examine your liver section tomorrow.
Complete part two of the packet.
Part One:
1. List the five steps of the scientific method.
63
a.
State the problem
b.
Research the problem
c.
Form a hypothesis
d.
Perform a controlled experiment
e.
Form a conclusion
2. What is the purpose of your experiment?
To determine if liver can be decellularized.
3. Name at least three things you have done to learn about (research) your problem.
Watched video clips about regenerative medicine, read a journal article
about liver decellularization, learned about liver anatomy.
4. What is your hypothesis? (best written as an “if/then” statement)
If a liver section is placed in a Triton X solution, then it will become
decellularized.
5. Identify the following for your experiment:
a. independent variable
Triton X % or SDS solution
b. dependent variable
Amount of decellularization
c. control (group)
Untreated (native) liver
d. experimental (group)
Liver sections that went into the flask w/ Triton X
e. constants
temperature, time, type of liver
6. Using detail and colored pencils, draw a picture of your liver before being placed into
the flask.
64
Part Two:
1. Retrieve your section of liver.
2. Using detail and colored pencils, draw what your liver section looks like after
treatment.
3. Describe at least three differences in your liver section between yesterday and today.
There should be a color change, few/no cells remaining, less mass, less volume.
4. If your experiment worked, what material are you looking at now? What is the main
component of that material?
You are left with extracellular matrix, which is mostly collagen.
5. Are you able to see any vascular tissue? Why is it important to keep the vascular
tissue when bioengineering a liver?
Depends on specimen. The vascular tissue will provide a way for new cells to get
65
nutrients and oxygen.
6. List at least three other possible independent variables that could be investigated in
terms of liver decellularization.
Concentration of solutions, temperature, time in solution, add an enzyme,
type of liver.
7. Does you data support your hypothesis? Explain.
Depends on results. However, the student needs to specifically state comparing
their results with the control.
8. How many specimen were in you experimental group? What is the minimum number
you need for a scientific experiment?
Depends on the class. At least three.
9. The next step to bioengineer a liver is to add stem cells. What type of stem cells
would you use, and explain why that is the best choice.
Answers can vary, but the most logical is the induced pluripotent stem cells
from the person receiving the new liver. These are less likely to be rejected,
are less likely to cause teratomas and are less controversial.
10. Give an argument as to how confident are you that researchers will be able to
bioengineer working organs, like livers, in the next 5 years. You must write a complete
paragraph, with at least three logical supporting statements based on what you have
learned during this unit.
Answers can vary, but the supporting statements need to make sense.
Researchers are optimistic that some working organs can be bioengineered
within five years.
66
Name___________________
Bioengineering a Liver
Now that you have learned about regenerative medicine, the role of the liver
and diseases of the liver, it is time to take the first step in making a new liver. This is
called decellularization, which strips away all the cells leaving only behind the
extracellular matrix which is then used as a scaffold to make the new liver.
Materials:
fresh chicken liver
flask
stir plate
SDS solution
camera
ruler
scalpel
stir bar
Triton X solution
colored pencils
dissection tray
Procedure:
1.
2.
3.
4.
5.
6.
7.
8.
Get a fresh chicken liver from your teacher.
Take several pictures of the liver.
Use the scalpel to cut a piece of liver no larger than 1 cm x 1 cm. For best
results, use a section where there are veins and arteries.
Choose which flask to put your liver piece into.
Complete part one of the packet while you wait.
After 40 minutes, empty the solution and put 500 mL of new solution into the
flask.
Examine your liver section tomorrow.
Complete part two of the packet.
Part One:
1. List the five steps of the scientific method.
67
a.
______________________________________________
b.
______________________________________________
c.
______________________________________________
d.
______________________________________________
e.
______________________________________________
2. What is the purpose of your experiment?
3. Name at least three things you have done to learn about (research) your problem.
4. What is your hypothesis? (best written as an “if/then” statement)
5. Identify the following for your experiment:
a. independent variable
_______________________________
b. dependent variable
_______________________________
c. control (group)
_______________________________
d. experimental (group)
_______________________________
e. constants
_______________________________
6. Using detail and colored pencils, draw a picture of your liver before being placed into
the flask.
68
Part Two:
1. Retrieve your section of liver.
2. Using detail and colored pencils, draw what your liver section looks like after
treatment.
3. Describe at least three differences in your liver section between yesterday and today.
4. If your experiment worked, what material are you looking at now? What is the main
component of that material?
69
5. Are you able to see any vascular tissue? Why is it important to keep the vascular
tissue when bioengineering a liver?
6. List at least three other possible independent variables that could be investigated in
terms of liver decellularization.
7. Does you data support your hypothesis? Explain.
8. How many specimen were in you experimental group? What is the minimum number
you need for a scientific experiment?
9. The next step to bioengineer a liver is to add stem cells. What type of stem cells
would you use, and explain why that is the best choice.
10. Give an argument as to how confident are you that researchers will be able to
bioengineer working organs, like livers, in the next 5 years. You must write a complete
paragraph, with at least three logical supporting statements based on what you have
learned during this unit.
70
LESSON SIX
TITLE: We Need Stem Cells
KEY QUESTION(S): What are the different types of stem cells? Why are stem cells important? Why are
stem cells controversial?
SCIENCE CONCEPTS: Cellular reproduction, differentiation.
OVERALL TIME ESTIMATE: One 50 minute class period.
LEARNING STYLES: Visual, auditory and kinesthetic.
VOCABULARY:
Adult stem cell: Stem cells taken from umbilical cord blood, bone marrow, or some organs.
Differentiation: Process an unspecialized cell takes to become specialized, changes form and function.
Embryonic stem cell: Stem cell taken from an embryo.
Growth factor: Proteins that turn genes on /off to promote specialization.
Induced pluripotent stem cell: An adult cell that has undergone a process to turn it into a stem cell.
In vitro fertilization (IVF): Combining sex cells (sperm/egg) in the laboratory to create embryos.
Zygote: The first stage of development when the sperm joins the egg.
LESSON SUMMARY: Students will read about the different types of stem cells and complete a
worksheet. They will then conduct a stem cell simulation to determine which types are better for use in
regenerative medicine.
STUDENT LEARNING OBJECTIVES WITH NEXT GENERATION SUNSHINE STATE STANDARDS:
1. Compare the different types of stem cells.
2. Explain which type of stem cell works best for regenerative medicine.
71
SC.912.L.1.1
Describe the scientific theory of cells (cell theory) and relate the history of its
discovery to the process of science.
SC.912.L.1.2
Relate structure to function for the components of plant and animal cells.
Explain the role of cell membranes as a highly selective barrier (passive and
active transport).
SC.912.L.14.6
Explain the significance of genetic factors, environmental factors, and
pathogenic agents to health from the perspectives of both individual and public
health.
MATERIALS:
ESSENTIAL:
 One lab packet per group of STO-120 Stem Cells from Science Take-out. All supplies are
included in the packet.
 One copy of the student worksheet in the packet, per student.
SUPPLEMENTAL:
BACKGROUND INFORMATION:
Once the liver scaffold is made out of the ECM, it must be seeded with stem cells. This activity attempts
to illustrate the various types of stem cells and the range of difficulty in getting them to differentiate.
While embryonic stem cells are easier to differentiate, they do come with some severe problems.
Embryonic stem cells are taken from embryos, which destroys the embryo. For many people this is
unethical. Another problem is that embryonic stem cells can form nasty tumors called teratomas. To
avoid this, researchers prefer to use stem cells from the patient’s own bone marrow or liver. This
activity from Science Take-out was probably written before the publication of induced pluripotent stem
cells (iPS cells). In 2007, Dr. Shinya Yamanaka was able to take a regular cell and change it into a stem
cell. There is a really good 15 minute video clip listed in the extension section that explains the induced
pluripotent stem cell procedure and can be shown to students.
ADVANCE PREPARATION:
Since this simulation comes pre-packaged, the teacher should go through a trial run before the lab.
PROCEDURE AND DISCUSSION QUESTIONS WITH TIME ESTIMATES:
See attached PDF from Science Take-out.
ASSESSMENT SUGGESTIONS:
For objective 1, student answers on questions #4-6 (part A) are the assessment.
For objective 2, student answer on question #5 (part B) is the assessment.
72
EXTENSIONS:
ACTIVITIES:
 Have students watch a 17 minute video clip explaining how Dr. Shinya Yamanaka
developed the process for induced pluripotent stem cells at:
www.youtube.com/watch?v=Q9-4SMGiKnE
 Have students complete an interactive tutorial from the University of Utah’s Genetic
Science Learning Center, which defines the different types of stem cells (adult,
embryonic, IPS), and their applications at:
http://learn.genetics.utah.edu/content/tech/stemcells/
LITERATURE: None
RESOURCES/REFERENCES:
73
LESSON SEVEN
TITLE: Bioengineer Me a Liver!: Part Two
KEY QUESTION(S): How do you decellularize a liver?
SCIENCE CONCEPTS: Experimental design, analyzing results.
OVERALL TIME ESTIMATE: One 50 minute class period for part two.
LEARNING STYLES: Visual, auditory, and kinesthetic.
VOCABULARY:
Constants: Factors that are kept the same throughout the experiment.
Control: Group that is not treated with the independent variable. Used to measure results against.
Dependent Variable: The change you are looking for, what you are measuring.
Experimental Group: Group that is treated with the independent variable.
Hypothesis: Prediction for the results of the experiment, best written as an “if-then” statement and
based on something measureable.
SDS: Sodium dodecyl sulfate, a detergent used to cause cell lysis.
Triton X 100: A non-ionic detergent used to cause cell lysis.
LESSON SUMMARY: Students will examine the results of their controlled experiment to decellularize a
chicken liver.
STUDENT LEARNING OBJECTIVES WITH NEXT GENERATION SUNSHINE STATE STANDARDS:
1. Compare experimental results with a control.
2. Determine if results support the hypothesis.
SC.912.N.1.1
74
Define a problem based on a specific body of knowledge, for example: biology,
chemistry, physics, and earth/space science, and do the following:
45. pose questions about the natural world,
46. conduct systematic observations,
47. examine books and other sources of information to see what is already
known,
48. review what is known in light of empirical evidence,
49. plan investigations,
50. use tools to gather, analyze, and interpret data (this includes the use of
measurement in metric and other systems, and also the generation and
interpretation of graphical representations of data, including data tables
and graphs),
51. pose answers, explanations, or descriptions of events,
52. generate explanations that explicate or describe natural phenomena
(inferences),
53. use appropriate evidence and reasoning to justify these explanations to
others,
54. communicate results of scientific investigations, and
55. evaluate the merits of the explanations produced by others.
SC.912.N.1.2
Describe and explain what characterizes science and its methods.
SC.912.N.1.3
Recognize that the strength of usefulness of a scientific claim is evaluated
through scientific argumentation, which depends on critical and logical thinking,
and the active consideration of alternative scientific explanations to explain the
data presented.
SC.912.N.1.5
Describe and provide examples of how similar investigations conducted in many
parts of the world result in the same outcome.
SC.912.N.1.6
Describe how scientific inferences are drawn from scientific observations and
provide examples from the content being studied.
SC.912.N.1.7
Recognize the role of creativity in constructing scientific questions, methods and
explanations.
MATERIALS:
ESSENTIAL:
 Treated chicken liver sections, one per lab group. While preferred, the treated section
may not be the same section that group prepared in Lesson Five/Day One.
 Beaker, one per lab group.
 Lab packet from Lesson Five/Day One, one per student.
SUPPLEMENTAL:
 Dissection microscopes to observe treated liver sections.
 Hand lens to observe treated liver sections, one per group.
75
BACKGROUND INFORMATION:
Liver sections have been treated with Triton X 100 1% solution for two hours and SDS .1% solution for
24-48 hours at this point. If the sections match the pictures at the end of Lesson Five, the process can
be stopped at 24 hours. If the sections are still brown, run the treatment for another 24 hours. What
remains is the extracellular matrix (ECM) which is mostly collagen. This ECM could then be seeded with
stem cells, ideally from the person who needs the new liver, which will grow on the scaffold and become
a new, functioning liver available for transplant.
Pictures of results of my experiment are posted at the end of this lesson.
ADVANCE PREPARATION:
The advanced preparation is to monitor the status of the liver sections when they are in the solutions,
checking for the end result of clear color. Leaving the sections in too long won’t ruin the experiment,
but they can get torn up by the stir bar and won’t look like the original piece much.
PROCEDURE AND DISCUSSION QUESTIONS WITH TIME ESTIMATES:
Students will need to get a section of treated chicken liver. It is best to put the section into a small
beaker with either SDS solution from the flask or water. This is because the treated liver won’t hold its
shape at this point, kind of a blob if placed directly onto a dissection pan. If available, students might
see more detail viewing the sections under a dissection microscope or hand lens, although clear
differences are readily apparent with the naked eye. The teacher should assist students transferring the
liver sections from the flask and into a beaker. This will take 5 minutes. Students are to then complete
part two of the lab packet. This should take 35 minutes followed by 5 minutes to clean up. With the
remaining time, ask students to read their answers for question #10. Have the class evaluate which
arguments are most convincing with supporting statements.
ASSESSMENT SUGGESTIONS:
For objective 1, student answers to question #2 and #3 are the assessment.
For objective 2, students answers to questions #10 is the assessment.
EXTENSIONS:
ACTIVITIES:
76

Several restaurants sell fried chicken livers. These could be served after the lab has
been completed.
 If there is a nearby hospital or histology lab, it might be possible to get some samples
processed. What you want to ask for is H &E staining (Hematoxylin and Eosin). This is a
standard procedure and is inexpensive. You would only need to have a couple of
samples/slides processed. The chemicals involved in the staining process make it unsafe
to do in a high school lab. It is best to have samples of the control liver and treated liver
for comparison. Slides should show a lack of cellular material in the treated sample. A
better staining technique is Masson’s Trichrome stain, which has collagen material
showing up as blue.
 Samples can be preserved in a NBF (neutral buffered phosphorus) or 60% ethanol into a
PBS (phosphate buffered saline) solution.
LITERATURE:
RESOURCES/REFERENCES:
Sullivan, D, Mirmalek-Sani, S, Deegan, B. Decellularization methods of porcine kidneys for whole organ
engineering using a high-throughput system. Biomaterials 2012; 33:7756-7764.
How to perform H&E Staining, University of Pennsylvania.
www.med.upenn.edu/mcrc/histology_core/documents/H_E_staining.doc , accessed 6/26/2013.
H&E Staining, www.youtube.com/watch?v=2D0rj0m6dVs , accessed 6/26/2013.
Masson Trichrome Staining, www.bcm.edu/rosenlab/?PMID=12997 , accessed 6/26/2013.
Masson’s Trichrome Stain, en.wikipedia.org/wiki/Masson’s_trichrome_stain , accessed 6/26/2013.
77
Untreated chicken liver sections
Liver starting to turn clear in Triton X 100
78
Decellularized chicken liver sections after 24 hours
Note the blood vessels still intact
79
Bioengineered Livers in the Classroom,
You can’t Really do that?
Post Test
This is only a pre-test to determine how much you know before starting the unit.
Place the letter of the best choice on the line.
_____ 1.
After decellularization, what remains? A) extracellular matrix
B) cytoplasm C) cell nuclei D) cytoplasm
_____ 2.
Which chemical, made by the pancreas, is used to regulate blood glucose
levels? A) pancreatic juice B) insulin C) glycogen D) collagen
_____ 3.
Juvenile diabetes is another name for? A) Type 1 diabetes B) Type 2
diabetes C) gestational diabetes D) prediabetes
_____ 4.
Of those with diabetes, about ____% have Type 1. A) 90% B) 50%
C) 25% D) 5%
_____ 5.
Which of the following is NOT a risk factor for diabetes? A) diet
B) exercise C) education level D) genetics
_____ 6.
What is the purpose of organ decellularization? A) remove only the bad
cells B) provide a scaffold for reseeding C) provide a means of gene
therapy D) reduce organ failure in diseased individuals
_____ 7.
Which of the following is NOT a complication of diabetes? A) limb
amputation B) blindness C) increased heart disease D) increased
Alzheimer’s
_____ 8.
What is the advantage of using embryonic stem cells? A) they don’t form
teratomas B) they only become one type of cell C) they are pluripotent
D) they are not controversial
_____ 9.
Which of the following is NOT a symptom of diabetes? A) loss of
appetite B) frequent urination C) sores that heal slowly D) excessive
thirst
_____ 10.
Normal blood glucose levels are about _____ mg/dL. A) 30 B) 100
C) 200 D) 300
_____ 11.
How did researchers conclude that scaffolding was safe? A) it passed
OSHA guidelines B) cells grew on it C) it failed to create an immune
80
response D) macrophages were able to destroy it
Use the following scenario to answer questions 12-16.
Dr. Sullivan is testing an implant to determine if it can be safely placed into the
body. He has treated the implants with different types of surfactants. The implant is
surgically placed into some rats, while other rats get surgery with no implant. After 8
weeks, he looks at the levels of inflammation associated at the implant site.
_____ 12.
The amount of inflammation would be the _______. A) independent
variable B) dependent variable C) control D) constant
_____ 13.
The type of rat used would be the _____. A) independent variable
B) dependent variable C) control D) constant
_____ 14.
The type of surfactant used to treat the implant would be the _____.
A) independent variable B) dependent variable C) control D) constant
_____ 15.
The rats that received surgery but no implants would be the _____.
A) independent variable B) dependent variable C) control D) constant
_____ 16.
What is the minimum number of trials or specimens for a scientific
experiment? A) 1 B) 2 C) 3 D) 100
_____ 17.
What is the biggest disadvantage of using embryonic stem cells?
A) they can form teratomas B) they are not pluripotent C) they only
work in the original host D) there are no embryos available to get stem
cells from
_____ 18.
What happens in Type 1 diabetes? A) glucose levels drop too low
B) collagen levels are too high C) no insulin is produced D) insulin is not
used by the cells
_____ 19.
What happens in Type 2 diabetes? A) glucose levels drop too low
B) collagen levels are too high C) no insulin is produced D) insulin is not
used by the cells
_____ 20.
When would someone get gestational diabetes? A) by not exercising
B) when pregnant C) after eating a low calorie diet D) after eating a high
cholesterol diet
_____ 21.
Of those with diabetes, about ____% have Type 2. A) 90% B) 50%
C) 25% D) 5%
81
_____ 22.
Adult onset diabetes is another name for? A) Type 1 diabetes B) Type 2
diabetes C) gestational diabetes D) prediabetes
_____ 23.
How many Americans are estimated to have prediabetes? A) 1 million
B) 10 million C) 50 million D) 80 million
_____ 24.
rate
What is the best prevention for Type 1 diabetes? A) reduce your heart
by exercising less B) eat a healthier diet C) increase the amount of
fructose in the diet D) there is no prevention
_____ 25.
rate
What is the best prevention for Type 2 diabetes? A) reduce your heart
by exercising less B) eat a healthier diet C) increase the amount of
fructose in the diet D) there is no prevention
_____ 26.
How many Americans have some form of diabetes? A) 20 million
B) 30 million C) 40 million D) 50 million
_____ 27.
What is the main type of cell found in the liver? A) hepatocyte
B) macrophage C) neurofibril D) thrombocyte
_____ 28.
Which of the following is NOT a function of the liver? A) metabolizes
drugs/medicine B) stores glycogen C) digests proteins D) metabolizes
lipids
_____ 29.
Which of the following diseases is NOT associated with the liver?
A) hepatitis B) malaria C) cirrhosis D) ischemia
_____ 30.
What is the purpose of adding a detergent/surfactant to an organ during
decellularization? A) strengthen the ECM B) cause cell lysis
C) keep vascular tissue intact D) maintain viability for transplant
82
Bioengineered Livers in the Classroom,
You can’t Really do that?
Pre/Post Test Answer Key
1.
2.
3.
4.
5.
6.
7.
8.
9.
10.
11.
12.
13.
14.
15.
83
A
B
A
D
C
B
D
C
A
B
C
B
D
A
C
16.
17.
18.
19.
20.
21.
22.
23.
24.
25.
26.
27.
28.
29.
30.
C
A
C
D
B
A
B
D
D
B
A
A
C
D
B