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Stem Cell Res. 2018 Oct;32:126-134. doi: 10.1016/j.scr.2018.09.006. Epub 2018 Sep 26.

Rapid and efficient differentiation of functional motor neurons from human iPSC for neural injury modelling.

Author information

1
Institute of Biomedical Engineering, Department of Engineering Science, University of Oxford, OX3 7DQ, UK.
2
Department Physiology, Anatomy and Genetics, University of Oxford, OX1 3QX, UK.
3
Department of Engineering Science, University of Oxford, OX1 3PJ, UK.
4
Institute of Biomedical Engineering, Department of Engineering Science, University of Oxford, OX3 7DQ, UK. Electronic address: hua.ye@eng.ox.ac.uk.

Abstract

Primary rodent neurons and immortalised cell lines have overwhelmingly been used for in vitro studies of traumatic injury to peripheral and central neurons, but have some limitations of physiological accuracy. Motor neurons (MN) derived from human induced pluripotent stem cells (iPSCs) enable the generation of cell models with features relevant to human physiology. To facilitate this, it is desirable that MN protocols both rapidly and efficiently differentiate human iPSCs into electrophysiologically active MNs. In this study, we present a simple, rapid protocol for differentiation of human iPSCs into functional spinal (lower) MNs, involving only adherent culture and use of small molecules for directed differentiation, with the ultimate aim of rapid production of electrophysiologically functional cells for short-term neural injury experiments. We show successful differentiation in two unrelated iPSC lines, by quantifying neural-specific marker expression, and by evaluating cell functionality at different maturation stages by calcium imaging and patch clamping. Differentiated neurons were shown to be electrophysiologically altered by uniaxial mechanical deformation. Spontaneous network activity decreased with applied stretch, indicating aberrant network connectivity. These results demonstrate the feasibility of this rapid, simple protocol for differentiating iPSC-derived MNs, suitable for in vitro neural injury studies focussing on electrophysiological alterations caused by mechanical deformation or trauma.

KEYWORDS:

Electrophysiology; Induced pluripotent stem cells (iPSC); Motor neuron differentiation; Neural injury

PMID:
30278374
DOI:
10.1016/j.scr.2018.09.006
[Indexed for MEDLINE]
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