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terms.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.This work was supported by the Netherlands Organisation
for Scientific Research (NWO), Rubicon Fellowship
(019.163LW.032) (to MvdH); the Brain Foundation, the
Walker Family, and the Perpetual Impact Philanthropy (grant
IPAP2017/0717) (to CB); the G. Harold & Leila Y. Mathers
Charitable Foundation, JPB Foundation, and the NIH (Grants
MH095741, MH092758, and U01 MH106882) (to FG). GY
was supported by R01 grants MH107369, HD085902, and
AI095277 from the National Institute of Health and Seed Grant
BRFSG-2014-14 from the Brain Research Foundation
