Application NoteChannel:
INa, IK
Cells: hiPSC-derived neuronsTools:
Patchliner®

Peri.4U and Dopa.4U stem cell-derived
neurons recorded on Nanion´s Patchliner®
The electrophysiology team at Nanion Technologies GmbH, Munich.
hiPSC neurons kindly provided by Axiogenesis, AG.
SummaryResults

Human induced pluripotent stem cell-derived neurons

NaV and KV currents of a Peri.4U cell with thefor studying the mechanisms underlying neurologicaldiseases and drug development. Axiogenesis provides
a number of hiPSC-neurons including dopaminergic
neurons (Dopa.4U) and peripheral neurons (Peri.4U),
amongst others. These neurons have been used on
in-vitro systems such as multielectrode arrays (MEA),

corresponding IV plots from an average of 8 cells areshown in Figure 1. See Table 1 for average peak amplitudes. A Boltzmann equation was used to fit the NaV IV

curve and Vhalf = -19 mV. Figure 2 shows NaV and KV currents of a Dopa.4U cell with the corresponding IV plots
from an average of 5 and 4 cells, respectively.
AB
0.4

immunocytochemistry and calcium imaging. They are0.2Voltage (mV)

an interesting model for studying neurological diseases
such as Parkinson’s Disease, as well as for efficacy, drug

-80

discovery and toxicity studies.

-40 -0.2-0.4

Dopa.4U and Peri.4U neurons in voltage and current-0.8

0.2 nA

clamp modes. The cell harvesting procedure was

-1.0

5 ms

1.0

optimized (using papain) to ensure that the cells retained

0.8

DCI/Imax

maintain ion channel expression present in these regions.
Due to their irregular shape (presence of processes),
success rate (typically 20 - 50% for RSeal >200 MΩ) was
lower than other cell types which have a smooth, round

Action potentials (AP) were also recorded and block of
the AP of Dopa.4U cells by lidocaine is shown.
CellRSeal(MΩ)

NaV peak (pA)

KV peak (pA)

Peri.4U

860 ± 101 (31)586 ± 135 (8)748 ± 191 (5)Dopa.4U

852 ± 477 (6)851 ± 300 (5)555 138 (4)Table 1: Seal resistance (RSeal) and peak amplitude values for NaV andKV recorded from Peri.4U and Dopa.4U cells recorded on the Patchliner®.
Number of cells shown in brackets.

Download more Application Notes from www.nanion.de0.60.40.2

shape, e.g. standard cell lines. Voltage-gated Na (NaV)and K (KV) currents were recorded in both cell types.

40-0.6In this study, the Patchliner was used to record fromproximal dendrites and initial axon segments in order toI/Imax(hiPSC-neurons) may provide a viable cellular model0.2 nA-80
40 ms-40
40
Voltage (mV)

Figure 1: A Raw NaV current traces from an exemplar Peri.4U neuron. B
Corresponding IV plot for an average of 8 cells. C Raw KV current tracesfrom an exemplar Peri.4U neuron. D Corresponding IV plot for an averageof 8 cells.

Application NoteA Boltzmann equation was used to fit the NaV IV curve

0 mVand Vhalf = -31mV.

B

0.2
Voltage (mV)-80
0.2 nA

-40

-0.2
-0.4

I/Imax

A40

-0.6

5 ms

-0.8
CControl
10 µM Lidocaine
Washout

20 mV
5 ms

Figure 3: AP elicited from a Dopa.4U neuron using a 1 ms pulse to 80 pAin control (black) and in the presence of 10 μM lidocaine (blue). The AP
could be recovered after washout of lidocaine (grey).

-1.0
1.0

In summary, Peri.4U and Dopa.4U neurons from Axiogenesis

0.8

to 50% (seal resistance > 200 MΩ). The harvesting protocol

I/ImaxD-75 mV

0.6
0.4
0.2

can be used on the Patchliner® with a success rate of up
using papain was developed in an attempt to retain thecell architecture at the expense of capture rate. It should
be noted, however, that the effect of detachment andloss of some neurite-located ion channels can change
the profile of cell currents, resting membrane potential

0.1 nA
100 ms-80-40
40Voltage (mV)

Figure 2: A Raw NaV current traces from an exemplar Dopa.4U neuron.
B Corresponding IV plot for an average of 5 cells. C Raw KV current
traces from an exemplar Dopa.4U neuron. D Corresponding IV plot for anaverage of 4 cells.

and responses to some neurotransmitters. Nevertheless,
NaV and KV currents were recorded in voltage clamp
mode and APs were elicited in current clamp mode.
This provides the opportunity to combine a cellular
neuronal model with higher throughput automatedelectrophysiology. In this way, such a cell model offers anFigure 3 shows an AP elicited from a Dopa.4U cell andalternative to primary neuronal cell cultures for studying

inhibition of the AP by lidocaine.

neuronal toxicity, disease research and drug discovery.

Methods

Patchliner®. For NaV currents, cells were stepped from aCells

increasing in 10 mV increments with each sweep up to

Human iPS cell-derived neurons (Peri.4UTM and Dopa.4UTM

60 mV. For KV currents, cell were stepped from a holding

holding potential of -100 mV to -80 mV for 20 ms and thenneurons) from Axiogenesis AG were used.

potential of -80 mV to -60 mV for 200ms and thenincreasing in 20 mV increments with each sweep up to 60

Cell culture

mV. For current clamp recordings, cells were sealed andCells were received as frozen aliquots and were plated

the whole cell configuration achieved in voltage-clamp

and cultured according to the manufacturer’s instruc-mode, following which, cells were switched to current

tions. Cells were harvested using papain.

clamp mode. Current was injected to maintain a constantmembrane potential of -75 mV (set individually for eachElectrophysiologycell) and action potentials were elicited using a 1 ms

Whole cell patch clamp recordings were conductedaccording to Nanion’s standard procedure for thecurrent pulse to the threshold required to elicit an action
potential (set individually for each cell).

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