Application NoteChannel:
Cells:

INa, IK, ICa, hERG
human iPS 		
cardiomyocytes
Patchliner®

Tools:

Voltage and current clamp recordings ofCor.4U® human iPS cell-derived cardiomyocytes on Nanion’s Patchliner®
The electrophysiology team at Nanion Technologies GmbH, Munich.
Cells kindly provided by Axiogenesis AG.

SummaryResults

Although mouse embryonic stem (ES) cell-derived cardi-

Figure 1 shows recordings in the voltage clamp mode ofomyocytes, e.g. Cor.At® cells from Axiogenesis, can pro-Cor.4U® cells exhibiting NaV (A) and KV (B) currents. An in-vide a useful model for drug discovery and safety test-

ward K+ current was observed when using an external so-

ing as an alternative to acutely dissociated rat or mouse

lution containing high K+ (C). This is confirmed to be hERG

cardiomyocytes, human induced pluripotent (iPS) cell-by the use of the specific antagonist E4031. In the current

derived cardiomyocytes have the potential to provide

clamp mode, spontaneous action potentials (AP) could

the ultimate model system for identifying potential anti-

be seen (D). Figure 2 shows both Na+ and Ca2+ currents asarrythmic effects of drugs during routine safety screening.

recorded in the voltage clamp mode. Furthermore, theAxiogenesis has now launched the Cor.4U human iPS

Ca2+ channel agonist BayK 8644 increases Ca2+ channel

cell-derived cardiomyocyte product line for use in test-

mediated currents.

ing the efficacy and safety of pharmaceutical therapies.

A

®

B

The ability to characterize the ion channel profile of thesecells and reliably record action potentials at a reasonable throughput is essential to fully realise the potentialof this kind of product line. Building on the success of Cor.

Patchliner® in the voltage and current clamp modes.
In this Application Note we present data using an 8-channel Patchliner® characterizing Cor.4U® cells. In the voltage
clamp mode, voltage-dependent Na+ (NaV), K+ (KV) andhERG (an inward current using a high K -containing ex+ternal solution) channel currents were recorded (Fig. 1).
When the Ca2+ channel agonist BayK 8644 was used a
voltage-gated Ca2+ (CaV) current could be recorded(Fig. 2). As expected, action potentials could be elicited in the current clamp mode (Fig. 3). The effect of thecompounds TTX and BayK8644 on the action potentials
evoked in Cor.4U cells is also shown (Fig. 3).

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50 ms

10 ms

cytes on the Patchliner®1,2, Cor.4U® human iPS cell-derived
cardiomyocytes have now been characterized on the400 pA

400 pAAt® mouse embryonic stem (ES) cell-derived cardiomyoC

Dcontrol
300 nM E4031
100 pA
100 ms

10 mV

100 msFigure 1:Currents recorded from Cor.4U® cells. A NaV current, B KV current and C
hERG inward current (recorded in the presence of high external K+) andblock by E4031. Spontaneous AP as recorded in the current clamp mode.

Application NoteA

BmV-40 -20

Caconjunction contribute to the action potentials which
are elicited in these cells. The compounds BayK 8644 and20 40

TTX modify the action potentials as expected.

-100

1 nANacardiomyocytes. These ion channels expressed in100pA

A-200

Control
10 µM BayK 8644

10 ms

40 mV
50 ms-95 mV

Figure 2:
A Raw data traces as recorded in the voltage clamp mode showing Na+and Ca2+ inward currents. B Ca2+ channel mediated currents in Cor.4U cells
could be increased using BayK 8644. IV plot in control conditions (closedcircles) and in the presence of BayK 8644 (open circles), raw data traces
not shown.

Figure 3 shows action potentials recorded from Cor.4U®

cells recorded in the current clamp mode in control
conditions (A) and in the presence of compounds BayK8644 and TTX (B, C). BayK 8644 elicits a lengthening of theduration of the AP (B) and TTX abolishes the AP (C). ThisBCcontrol
10 µM BayK 8644

40 mV-100 mV

control
10 µM TTX
washout
40 mV

20 ms

20 ms

Figure 3:
Action potentials recorded from Cor.4U® cells in the current clamp mode. AIn control conditions, B In the presence of BayK 8644 and C in the presenceof TTX.

effect is reversible upon washout of TTX.
The data provide evidence that the Patchliner® is well
In conclusion, Cor.4U®

cells exhibit the ion channel

suited to recording Cor.4U® cells in a higher throughput

profile (NaV, KV, CaV and hERG) expected in humanenvironment given it’s flexibility and reliability.

References

Electrophysiology
Whole cell recordings were conducted according to

1. Stoelzle et al., 2011. JBS. 16 (8): 910–916

Nanion’s standard procedure for the Patchliner®. Current-

2. Stoelzle at al., 2011. Front Pharmacol. 2 (76) doi: 10.3389/

voltage recordings were made using voltage steps fromfphar.2011.00076-60 mV to 50 mV for 20 ms increasing in 10 mV steps, froma holding potential of -120 mV (NaV); -60 mV to 60 mV forMethods

200 ms increasing in 20 mV steps from a holding potentialof -80 mV (KV); -40 mV to 50 mV for 100 ms increasing in 10Cells

mV steps from a holding potential of -100 mV (CaV; in theCor.4U® cells were supplied by Axiogenesis AG.

presence or absence of 10 µM BayK 8644). Inward hERG
current was elicited by stepping from the pre-pulse +40
mV to -120 mV using an external solution containing 126

Cell culture

mM K+. APs were elicited using a 1 ms current pulse to theCells were cultured and harvested according toNanion’s standard cell culture protocol for stem cell-de-threshold necessary to generate an action potential ineach cell (calculated for each cell individually).

rived cardiomyocytes.

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