Complementary automated patch clamp, extracellular field potential andimpedance recordings of iPSCs: safety screening tool box for the future
Elena Dragicevic1, Nadine Becker1, Sonja Stölzle-Feix1, Ulrich Thomas1, Krisztina Juhasz1, Leo Doerr1, Tom Goetze1, Markus Rapedius1,Daniel Millard2, Stacie Chvatal2, Anthony Nicolini2, Matthias Beckler1, Michael George1, Andrea Brüggemann1, Niels Fertig1
1Nanion Technologies GmbH, Gabrielenstr. 9, 80636 Munich, Germany

2Axion BioSystems, Inc., 1819 Peachtree Road, Suite 350, Atlanta, GA, 30309

Patchliner

CardioExcyte 96

Minimized Cell Usagein Automated Patch Clamp

Axion Maestro
Axion Multi-well MEA Technology

CardioExcyte 96 Sensor Technology

First-in-class high-throughput
MEA System with a powerful Software

Maestro with EC mini

CE 96

robotic pipette

Impedance Comprehensive Extracellular
Field
safety assays
Potential
Contractility

int. solution

electricalconduction(e.g. Long QTSyndrome)

12-well
64 electrodes/well
Combined Impedanceand EFP measurements

Cardiac
ActionPotential

catch rate (%)

Becker, N., et al., Minimized cell usage for stem cell-derived and primary cells on an
automated patch clamp system, Journal of Pharmacological and Toxicological Methods (2013)

10010010080

8080606060

CHO cells

40r² = 0.9420

y = 21.12ln(x) – 39.427

00200

400

600

iCell® Neurons

4020y = 27.593ln(x) – 92.001

00cells added per well

400

800

Cell Quality Control•

Toxicity Studies

Clinical ECG

Well Raster

Effect ofCompounds

CardiacMetrics
NeuronalMetrics

y = 18.678ln(x) – 45.98600cells added per well•r² = 0.77

201200Single SpikeRasterCor.4U®

40

r² = 0.91

Cell Proliferation Assays

Raw Data10 sCM-MEA
Field Potential•

96-well
8 electrodes/well

Cardiac and Neuronal Applications

Extended Application: adhesion and proliferation

chip hole

48-well
16 electrodes/well

100 mVcell suspension

Ion-channel
modulation:
Na+, Ca2+, K+,
hERG

Control(e.g.
inotropiceffects)

PTX

ext. solution

400

800

1200

cells added per wellcellattachment

Catch-rate using cell-lines and stem cell-derived cells. Only about 400 cells per recording
well are needed in order to perfom successfull automated patch-clamp measurements
using the Patchliner.

Action Potential Pharmacology ofCor.4U ® Cardiomyocytes

iCell® Cardiomyocytes, CDI
CM-MEA (Cardiomyocyte-Microelectrode Array)
bridges the gap between patch clamp-based
electrophysiology and clinical electrocardiograph(ECG) recording, bringing “heartbeats-in-a-dish” tothe benchtop.

cells start beating

synchronized beating
network of cells, monolayer

Comprehensive in vitro Proarrhythmia
Assay (CiPA)

CardioExcyte 96 Compound Profiling
Impedance

The synchronous firing pattern across a neuronal
network of cells over MEA provides a phenotype
for establishing “seizure-in-a-dish” models

EFP

Baseline

Induced action potentials

Spontaneous action potentials

DC

Dofetilide
100 nMcontrol
100nM Isoproterenol
washout

Dofetilide

20 mV

Continuous recording enables detectionof rare arrhythmic events

20 ms

L-Type Ca2+ channel activator Bay K
8644 prolongs action potentials

β-adrenergic agonist Isoproterenol
enhances action potentials

Voltage- and ligand gated currents
recorded in iCell® neurons

Block of Na+ current by increasing
concentrations of TTX (IC50 = 4.9 nM (n =1)).

Nifedipine

Axion’s software suite, AxIS, provides access to reliable measures of the four primary
endpoints of CiPA protocol: (1) Depolarization, (2) Repolarization, (3) Beat Timing, and (4)Arrhythmia. Additional measures, such as conduction velocity and pacer orientation, arealso computed to ensure data quality and enable further quantification of the functional
response to compound addition.

Activation of GABA currents andpartial block by bicuculline.

HTS Cardiac panel according to CiPA
Cardiac AP and currents

CardioExcyte 96 Environmental Control

CiPA pilot study on the Maestro Platform

Cardiac Panel Protocol
ms

SyncroPatch384PE421

Propagation Delay

30

iPSC Cardiomyocytes on MEA

recording

Cardiomyocyte beat activity

dosing“T-wave” detection

Beat propagation

analyisis window
minutes

Sager, P.T., et al., 2014.

CardioExcyte 96
Environmental Control•

Replaces the Incubator•

Minimal space requirements•

Easy assessment of
temperature-dependent
effects

Moxifloxacin

cellattachment

cells start beating

60300

Nifedipine

Mexiletine

synchronized beating
network of cells, monolayer

iCell® Cardiomyocytes², CDI• Temperature control up
to 40°C• 5 % CO2• 95 % humidity
Six different cardiac channels recorded simultaneously on the SyncroPatch® 384PE.
Screenshot shows data acquisition and analysis software performing recordings from HEKor CHO cells expressing the CiPA stipulated ion channels (ChanTest) activated within
one single experiment on the SyncroPatch® 384PE.

A snapshot of CiPA MEA pilot study results on the Maestro platform: top (red) and bottom(blue) row graphs reflect data from cell lines “A” and “B”, respectively, when treatedwith the compound indicated. Each of the 6 sites are independent testing locations.