Pharmacological Profiling of Purified Human Stem Cell-Derived and Primary Mouse Motor Neurons

Brian J. Wainger,Joan Koh,Kevin Zhu,Daniel Moakley,Eugene Berezovski,Joao D. Pereira,Daniel M. DuBreuil,Anna-Claire Devlin

doi:10.1038/s41598-019-47203-7

Abstract

Directed differentiation of human pluripotent stem cells (hPSCs) has enabled the generation of specific neuronal subtypes that approximate the intended primary mammalian cells on both the RNA and protein levels. These cells offer unique opportunities, including insights into mechanistic understanding of the early driving events in neurodegenerative disease, replacement of degenerating cell populations, and compound identification and evaluation in the context of precision medicine. However, whether the derived neurons indeed recapitulate the physiological features of the desired bona fide neuronal subgroups remains an unanswered question and one important for validating stem cell models as accurate functional representations of the primary cell types. Here, we purified both hPSC-derived and primary mouse spinal motor neurons in parallel and used extracellular multi-electrode array (MEA) recording to compare the pharmacological sensitivity of neuronal excitability and network function. We observed similar effects for most receptor and channel agonists and antagonists, supporting the consistency between human PSC-derived and mouse primary spinal motor neuron models from a physiological perspective.

Highlights

Deriving specific disease-relevant cellular subtypes from human pluripotent stem cells may serve potentially valuable roles in disease modelling, cell replacement, and drug development[1]
HB9 is an early marker of post-mitotic spinal motor neurons and is expressed in both human pluripotent stem cells (hPSCs)-derived and embryonic mouse spinal motor neurons[16]
To derive human motor neuron-enriched cultures, a hPSC line with a GFP expression marker driven by the HB9 promoter was differentiated using a monolayer strategy, and HB9:GFP-positive neurons were isolated using fluorescence-activated cell sorting (FACS)[3,17]

Summary

Introduction

Deriving specific disease-relevant cellular subtypes from human pluripotent stem cells (hPSCs) may serve potentially valuable roles in disease modelling, cell replacement, and drug development[1]. Focusing on physiology and the cohort of functional ion channels in specific neurons can provide a parallel functional assessment of the value of human stem cell-based models. Recent clinical phenotypes have raised the question as to whether abnormal neuronal excitability contributes to specific neurological diseases and whether the physiological features and functions of neuronal populations www.nature.com/scientificreports/. Www.nature.com/scientificreports may contribute to their selective vulnerabilities This possibility seems strong in familial epilepsies and rare pain syndromes due to mutations in specific sodium and potassium channels[9]. While a group of stem cell modelling studies have already documented abnormal motor neuron excitability in ALS, the underlying assumption that the physiology of hPSC-derived motor neurons accurately reflects the physiology of bona fide motor neurons has not been addressed. The physiological concordance of the model neurons with primary ones may be important in modelling diseases of specific vulnerable neuronal types

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