title: Characterizing human ion channels in induced pluripotent stem cell-derived neurons.
abstract: Neurons derived from human-induced pluripotent stem cells were characterized using manual and automated patch-clamp recordings. These cells expressed voltage-gated Na(+) (Na(v)), Ca(2+) (Ca(v)), and K(+) (K(v)) channels as expected from excitable cells. The Na(v) current was TTX sensitive, IC(50) = 12 ± 6 nM (n = 5). About 50% of the Ca(v) current was blocked by 10 µM of the L-type channel blocker nifedipine. Two populations of the K(v) channel were present in different proportions: an inactivating (A-type) and a noninactivating type. The A-type current was sensitive to 4-AP and TEA (IC(50) = 163 ± 93 µM; n = 3). Application of γ-aminobutyric acid (GABA) activated a current sensitive to the GABA(A) receptor antagonist bicuculline, IC(50) = 632 ± 149 nM (n = 5). In both devices, comparable action potentials were generated in the current clamp. With unbiased, automated patch clamp, about 40% of the cells expressed Na(v) currents, whereas visual guidance in manual patch clamp provided almost a 100% success rate of patching "excitable cells." These results show high potential for pluripotent stem cell-derived neurons as a useful model for drug discovery, in combination with automated patch-clamp recordings for high-throughput and high-quality drug assessments at human neuronal ion channels in their correct cellular background.
authors.affiliation: Nanion Technologies GmbH, Munich, Germany. ali@nanion.de
authors.forename: Alison
authors.initials: A
authors.lastname: Haythornthwaite
authors.forename: Sonja
authors.initials: S
authors.lastname: Stoelzle
authors.forename: Alexander
authors.initials: A
authors.lastname: Hasler
authors.forename: Andrea
authors.initials: A
authors.lastname: Kiss
authors.forename: Johannes
authors.initials: J
authors.lastname: Mosbacher
authors.forename: Michael
authors.initials: M
authors.lastname: George
authors.forename: Andrea
authors.initials: A
authors.lastname: Brüggemann
authors.forename: Niels
authors.initials: N
authors.lastname: Fertig
mesh_terms: D001640:Bicuculline / Q000494:pharmacology; D015220:Calcium Channels / Q000378:metabolism; D002454:Cell Differentiation; D055808:Drug Discovery / Q000379:methods; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology*; D020128:Inhibitory Concentration 50; D015640:Ion Channel Gating / Q000187:drug effects* / Q000502:physiology; D007473:Ion Channels / Q000378:metabolism*; D008564:Membrane Potentials / Q000187:drug effects / Q000502:physiology; D009474:Neurons / Q000187:drug effects / Q000378:metabolism*; D009543:Nifedipine / Q000494:pharmacology; D018408:Patch-Clamp Techniques / Q000379:methods*; D024642:Potassium Channels, Voltage-Gated / Q000378:metabolism; D013779:Tetrodotoxin / Q000494:pharmacology; D061566:Voltage-Gated Sodium Channels / Q000378:metabolism; D005680:gamma-Aminobutyric Acid / Q000494:pharmacology
chemical_list: D015220:Calcium Channels; D007473:Ion Channels; D024642:Potassium Channels, Voltage-Gated; D061566:Voltage-Gated Sodium Channels; D013779:Tetrodotoxin; D005680:gamma-Aminobutyric Acid; D009543:Nifedipine; D001640:Bicuculline