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Title: Patch-Seq Protocol to Analyze the Electrophysiology, Morphology and Transcriptome of Whole Single Neurons Derived From Human Pluripotent Stem Cells
Authors: van den Hurk, M.
Erwin, J.A.
Yeo, G.W. 
Gage, F.H.
Bardy, C.
Keywords: Cellular phenotyping
Human neuron transcriptome
Induced pluripotent stem cell (iPSC)
Neuronal diversity
Patch clamping
Single-cell RNA-seq
Issue Date: 2018
Publisher: Frontiers Media S.A.
Citation: van den Hurk, M., Erwin, J.A., Yeo, G.W., Gage, F.H., Bardy, C. (2018). Patch-Seq Protocol to Analyze the Electrophysiology, Morphology and Transcriptome of Whole Single Neurons Derived From Human Pluripotent Stem Cells. Frontiers in Molecular Neuroscience 11 : 261. ScholarBank@NUS Repository.
Rights: Attribution 4.0 International
Abstract: The human brain is composed of a complex assembly of about 171 billion heterogeneous cellular units (86 billion neurons and 85 billion non-neuronal glia cells). A comprehensive description of brain cells is necessary to understand the nervous system in health and disease. Recently, advances in genomics have permitted the accurate analysis of the full transcriptome of single cells (scRNA-seq). We have built upon such technical progress to combine scRNA-seq with patch-clamping electrophysiological recording and morphological analysis of single human neurons in vitro. This new powerful method, referred to as Patch-seq, enables a thorough, multimodal profiling of neurons and permits us to expose the links between functional properties, morphology, and gene expression. Here, we present a detailed Patch-seq protocol for isolating single neurons from in vitro neuronal cultures. We have validated the Patch-seq whole-transcriptome profiling method with human neurons generated from embryonic and induced pluripotent stem cells (ESCs/iPSCs) derived from healthy subjects, but the procedure may be applied to any kind of cell type in vitro. Patch-seq may be used on neurons in vitro to profile cell types and states in depth to unravel the human molecular basis of neuronal diversity and investigate the cellular mechanisms underlying brain disorders. © 2018 van den Hurk, Erwin, Yeo, Gage and Bardy.
Source Title: Frontiers in Molecular Neuroscience
ISSN: 1662-5099
DOI: 10.3389/fnmol.2018.00261
Rights: Attribution 4.0 International
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