Application of Human iPSC-derived Cardiomyocytes
and Neurons Using the MEA Technology
Giorgia Salvagiotto, Sabine Lange, Martin Augustin, Kile Mangan, Coby Carlson,
Arne Thompson, Susan DeLaura and Blake Anson.
Target
Identification
Target
Validation
Compound
Screening
Lead
Optimization
Preclinical
Trials
Cellular Dynamics International, Madison, WI USA
Clinical
Trials
iCell Cardiomyocytes show native
biology and function
Abstract
Multielectrode Array (MEA) technology
is a non-invasive, label-free platform
that measures the electrical activity of
single cells or cellular network at
medium to high throughput level. In
particular, measurements of local field
potentials
of
cardiomyocytes
monolayers allow the assessment of the
effect of cardioactive compounds on
cardiac electrophysiology, whereas
extracellular single-unit recordings of
neuronal
cultures
enable
the
investigation
of
neurotransmission
modulators on neuronal network activity.
Whole-Genome
Gene Expression
iCell Neurons show native
biology and function
Protein Expression
Gene Expression
Protein Expression
-------Myocardium
____ PromoCell
____ Celprogen
------ Adult myocardium
____ d28 iCell CM
____ d120 iCell CM
Here we present iCell® Cardiomyocytes
and iCell Neurons:
 Highly pure (>95%) human cells
 Industrialized manufacture at high
volumes and high quality
 Cryopreserved, ready to use
 Native genotypic, phenotypic, and
functional characteristics.
Relevant & temporally
stable in culture
Exhibit normal
cardiomyocyte markers
Babiarz et al., 2012
Kattman et al., 2011
Exhibit expected
neuronal markers
Xu et al., 2011
Electrophysiology, E-C
Coupling, Contractility
Metabolism
Validated forebrain
identity
Electrophysiology
Dage et al., 2014
20
Ca2+ Transient
10
HG= High Glucose
LG = Low Glucose
FA = Fatty Acid
GAL = Galactose
0
0mV
NMDA S/B= 1.4
Glutamate S/B =3
-10
Neurite Outgrowth
-20
10mV
1sec
-30
-40
-50
TuJ 1
DAPI
Example application data of the
functional utility of iCell Cardiomyocytes
and iCell Neurons using the MEA
technology will be presented, illustrating
how these cellular models have been
used for toxicity studies, are currently
evaluated for preclinical toxicity testing
and disease modeling, and are now
creating
new
opportunities
for
therapeutic decision-making.
Substrate dependent, active
mitochondria
Control
Enables mechanistic toxicity
testing
Ma et al., 2011, Puppala et al., 2012
Rana et al., 2012
-60
BDNF
Spontaneous neuronal
network behavior
Enables mechanistic toxicity
testing
For additional data:
Haythornthwaite et al., 2012
iPSC-derived Cardiomyocytes and MEA: a reliable assay for toxicity testing and disease modeling
iCell Cardiomyocytes monolayers reflect organotypic behavior
iCell Cardiomyocytes detect potentially fatal
compounds cheaper and faster
The iPSC technology enables access
to healthy and diseased human biology
Control
Control
Control
30 nM Cisapride
100 nM Astemizole
LQT2
Pharmacological responses of iCell Cardiomyocytes to drug-induced electrophysiological perturbations can be
assessed using the MEA technology. Treatment with E4031 (IKr channel blocker) and Nifedipine (Ica-L channel blocker)
prolongs and shortens the field potential duration (FDP), respectively (Harris et al., 2013).
Astemizole (anti-histamine) and Cisapride (acid reflux disease)
were withdrawn from the market in 1999 and 2000, respectively,
due to long QT syndrome and arrhythmias. The phenotypes
could be detected on iCell Cardiomyocytes using MEA.
F
P
D
2000
1000
m
s
1500
(
)
H
z
500
)
iCell Cardiomyocytes responses match those of
more labor-intensive in vitro assays (rabbit wedge)
MEA Frequency
F
r
e
q
u
e
n
c
y
2500
(
CellTiter-Glo®
Telemetry reading of a RO5657-treated monkey
MEA FPD
LQT2
10 µM RO5657
RO5657 arrhythmia was not detected until non-human primate
studies, whereas iCell Cardiomyocytes detected the toxicity
during in-vitro studies (Misner et al., 2012, Guo et al., 2011).
iPSC-derived Neurons and MEA: a real-time assay of network activity
iCell Neurons culture reflect neuronal network behavior
Activity regulators
A
B
CDI’s new MEA analysis tool: the
MATLAB-based application called
iCell NeuroAnalyzer.
Control
1. Mean Firing Rate –
Influence on Inhibition (IoI)
2. Bursting Rate –
Influence on Excitation (IoE)
3. Intensity within the Bursts –
Influence on Connectivity (IoC)
GABAzine
C
Control
CellTiter-Glo®
Synaptic stimulators
GABAzine
GABAzine
Representative data outputs from iCell Neurons
analyzed on the MEA system. A) Real-time heat map of
electrical activity across the plate; B) Action potentials and
C) Raster plots recorded from a single electrode before
and after treatment with the GABAA receptor inhibitor
GABAzine.
www.cellulardynamics.com
Pharmacological responses of iCell Neurons to druginduced network perturbations can be assessed using the
MEA technology and quantified using the iCell
NeuroAnalyzer. Treatment with neuronal activity regulators
and synaptic stimulators produced the expected effect on
the firing rate, bursting rate, and bursting intensity of the
neuronal network.
Madison, WI USA
0.5
0.4
0.3
0.2
0.1
LQT2
Control
Control
The iPSC technology enables access to
human tissues from healthy and clinical
genotypes from which different cell types can
be generated.
An LTQ2 patient iPSC line carrying an early
stop codon in the hERG coding region was
used to generate cardiomyocytes. The iPSCderived cardiomyocytes recapitulated the LTQ2
phenotype characterized by:
MEA reading of RO5657-treated iCell Cardiomyocytes
“MEA assays using iPSC-Cardiomyocytes offer a reliable,
cost effective surrogate to preclinical in vitro testing, in
addition to the 3Rs (refine, reduce, and replace animals in
research) benefit” (Harris et al., 2013).
0.6
0
0
Drug-induced effects on cardiac conduction
velocity can also be calculated on the MEA.
Treatment with Lidocaine (INa blocker) slows down
or delays the conduction velocity of iCell
Cardiomyocytes in a concentration-dependent
manner.
0.7
- Delayed repolarization
- Prolonged QT interval
- Polymorphic ventricular tachycardia
Summary
CDI is the leading producer of human iPS cellderived tissue types.
The data presented here highlight the utility of the
MEA system to gather relevant mechanistic data on
the
electrophysiological
activity
of
human
cardiomyocytes and neuronal network using iCell
Cardiomyocytes and iCell Neurons that:
 Recapitulate human behavior of the native cell
counterparts
 Provide a tool for retrospective investigative
toxicology
 Enable relevant, cost-effective, and highly
predictive preclinical testing
 Are produced through consistent and robust
manufacture
 Can be differentiated from healthy and
diseased iPSC lines, offering a tool for disease
in vitro modeling.
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