Author:
Le Cann Kim,Foerster Alec,Rösseler Corinna,Erickson Andelain,Hautvast Petra,Giesselmann Sebastian,Pensold Daniel,Kurth Ingo,Rothermel Markus,Mattis Virginia B.,Zimmer-Bensch Geraldine,von Hörsten Stephan,Denecke Bernd,Clarner Tim,Meents Jannis,Lampert Angelika
Abstract
AbstractHuntington’s disease (HD) is an autosomal dominant neurodegenerative disorder caused by an expanded polyglutamine repeat in the huntingtin gene. The neuropathology of HD is characterized by the decline of a specific neuronal population within the brain, the striatal medium spiny neurons (MSNs). The origins of this extreme vulnerability remain unknown. Human induced pluripotent stem cell (hiPS cell)-derived MSNs represent a powerful tool to study this genetic disease. However, the differentiation protocols published so far show a high heterogeneity of neuronal populations in vitro. Here, we compared two previously published protocols to obtain hiPS cell-derived striatal neurons from both healthy donors and HD patients. Patch-clamp experiments, immunostaining and RT-qPCR were performed to characterize the neurons in culture. While the neurons were mature enough to fire action potentials, a majority failed to express markers typical for MSNs. Voltage-clamp experiments on voltage-gated sodium (Nav) channels revealed a large variability between the two differentiation protocols. Action potential analysis did not reveal changes induced by the HD mutation. This study attempts to demonstrate the current challenges in reproducing data of previously published differentiation protocols and in generating hiPS cell-derived striatal MSNs to model a genetic neurodegenerative disorder in vitro.
Funder
Deutsche Forschungsgemeinschaft
Interdisciplinary Centre for Clinical Research within the faculty of Medicine at the RWTH Aachen University
RWTH Aachen
Publisher
Springer Science and Business Media LLC
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