Microsystems for Stem
Cell Research
Professor Pasi Kallio
Micro- and Nanosystems Research Group
Department of Automation Science and Engineering
BioMediTech
Tampere, Finland
Outline
• BioMediTech
• National ”Human Spare Parts” Programme
• Microsystems for Stem Cell Research
Institute Background
Joint institute of University of Tampere &
Tampere University of Technology,
i Tammerfors
BioMediTech created in 2011
• Over 250 scientists
• World-class basic and
translational research
Life Sciences and Medical Technology
Over 250 scientists conducting basic and
translational research in life sciences and medical
technology
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Tissue Engineering & Stem Cell Technology
Biomaterials
Sensor and Actuator Technologies
Measurement and Imaging Technologies
Biotechnology
Immunology
Cancer and Mitochondrial Research
Systems Biology
Bioinformatics
Computational Methods in Biomedicine
Kauppi & Hervanta Campuses
KAUPPI
BioMediTech
Outline
• BioMediTech
• National ”Human Spare Parts” Programme
• Microsystems for Stem Cell Research
Human Spare Parts Program
Combines expertise in
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biomaterials
microsystem and sensor technology
biomedical engineering
stem cells
Development of tissue engineering
products and supporting
technologies
Over 20 M€ project funding (2011-2015)
• Finnish Funding Agency for Technology and
Innovation (TEKES)
• Academy of Finland
• Council of Tampere Region
• EU
Human Spare Parts Program
HEART
NEURONS
RETINA
Cell models for
Blood-retinaCell models for
TECHNOLOGIES
(sensors,
actuators,
signals,
materials)
neurodegenerati
barrier
(BRB)
inherited heart
ve diseases
model for drug
diseases
development
BONE
Human Spare Parts - Technology
• Automated cell culture environments and
stimulation
• Prof. Pasi Kallio Group
• Sensor and measurement solutions
• Prof. Jukka Lekkala Group
• Imaging and image processing
• Prof. Jari Hyttinen Group
• Biomaterials
• Prof. Minna Kellomäki Group
Engineers?
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Cell models are not truly functional without an environment
which
• mimics body the conditions to keep the cells alive
• promotes cell functionalities, healthy and diseased
• mimics disease conditions
• allows drug exposure
• provides means to
characterize cell
responses
• provides support for
analysis
• ...
Vision: Autonomous Cell Stimulation and
Measurement Environment for Phenotypic Studies
Vision: Autonomous Cell Stimulation and
Measurement Environment for Phenotypic Studies
Outline
• BioMediTech
• National ”Human Spare Parts” Programme
• Microsystems for Stem Cell Research
Heart and Bone
• Cell mechanobiology using microsystems
Adj. Prof.
Susanna
Miettinen
Adj. Prof.
Katriina
Aalto-Setälä
Mechanical Stimulation of Stem Cells
• Objective and motivation
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To mechanically stimulate stem cells when
differentiating them to cardiac or osteogenic cells
In order to improve their functionality
Mechanical Stimulation of Stem Cells –
Our Approach
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Cells grow on a coated thin PDMS membrane which is
pneumatically deformed
Kreutzer et al. J. of
MedEng&Physics 2014
Computational Model and Optimization
using Comsol Multiphysics
Zhao et al. IEEE ICMA
2014
Coating
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PDMS needs coating for cells to grow
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Collagen protein is widely used
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Typically used coating methods are insufficient in cell stretching
à Collagen and cells break off
à Cells do not survive long experiments
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Covalent bonding needed
Cardiac Cell Experiments
Stretching: final strain amplitude
5%, frequency 1 Hz
Cardiac Cell Experiments –
Functionality Study
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Visual observation
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Beating cells with a proposed
stretching sequence
• Maturation analysis using quantitative real-time polymerase
chain reaction
– relative gene expression levels of stretched and unstretched
control samples were determined
– No significant difference with these parameters were found
– Uniaxial stretching with cardiac cells
AFM Study – Beating Force of Cardiac
Cell
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Measurement results
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Cardiomyocyte imaged by AFM
Liquid cell in AFM
Average beating rate 60-72 beats per minute
Typical beating force 2.5 - 7 nN
Typical beating amplitude 0.04-0.1 µm
Courtesy of Prof.
Jukka Lekkala, TUT
Bone: Mechanical Stimulation and
Characterization
Stretching structures
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Equiaxial and uniaxial
AFM study
Grooves for alignment
7
Apparent Young’s Modulus [kPa]
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0
Day 3
Static Day 13
Dynamic day 13
Brain
Dr. Susanna
Narkilahti
Characterization of Neural Cells
1. Gene expression analysis
4. Single cell electrophysiology (patch clamp)
1 3 6 9 12 15 wks
Oct-4
a-fp
T
Musashi
Pax-6
Nestin
Map-2
GFAP
Olig1
2. Protein expression analysis
NG2/GalC
MAP2/B-tubulin
5. Neuronal network analysis (Ca-imaging)
MAP2/NESTIN
DAPI/GFAP/Phalloidin
MAP2
3. Cell morphology analysis
6. Repeated neuronal network analysis (MEA)
Heikkilä et al., Exp Neurol 2009
MicroElectrode Arrays (MEAs)
Baseline
CNQX
CNQX+D-AP5
Washout
GABA
Washout
Bicuculline
Results similar to mouse embryonic stem cell derived neuronal networks (Illes et al, 2007)
Gas Supply Systems
Long-term MEA recordings outside an incubator (under
microscope)
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evaporation of medium
osmolarity changes
pH changes if no 5% CO2 present
Gas Supply Systems
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Structures from PDMS
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Biocompatible, optically good, transparent, gas permeable
Provides
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Humidity 95% ± 5%
5% CO2
Gas out
(1)
MEA
Gas in
Gas Supply Systems (CO2)
CO2 concentration in cell area [%]
gas ring
gas through
top PDMS
pH of culture media after 3days (without cells)
~7.4
Gravity-driven flow for Nutrient and
Drug Delivery
Modelling Drug Delivery
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Analytically solving time-dependent
inlet pressure (flow rate)
Shear stress [Pa]
Simulink model
p(t)
Concentration distribution
Mäki et al. 2014 FEDSM2014
Summary
• We have well-understood and characterized
solutions for
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Equiaxial and uniaxial cell stretching
Gas (CO2 and O2) supply
Perfusion and drug delivery
Steps towards Commercialization
DrugPermeA
SpikeBooster
RoundLabOut
Conclusions
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To get the cells beating or
signalling, you need a
devoted and
multidisciplinary team
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Cell biologists
Clinicians
Biomaterial engineer
Microfluidics engineer
Automation and
instrumentation engineer
Biosignal engineer …
Acknowledgement
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Collaborators
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Prof. Katriina Aalto-Setälä, UTA
Prof. Susanna Miettinen, UTA
Dr. Heli Skottman, UTA
Dr. Susanna Narkilahti, UTA
Prof. Markku Kulomaa, UTA
Prof. Timo Ylikomi, UTA
Dr. Vesa Hytönen, UTA
Prof. Matti Vilkko, TUT
Prof. Seppo Kuikka, TUT
Prof. Jukka Lekkala, TUT
Prof. Jari Hyttinen, TUT
Prof. Minna Kellomäki, TUT
Dr. Marja-Leena Linne, TUT
• Research Group at TUT
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Joose Kreutzer
Antti Mäki
Samu Hemmilä
Lassi Sukki
Feihu Zhao
Joni Leivo
Et al
• Funding
– Tekes
– The Academy of Finland
– Several Finnish companies
Thank You for Your Attention