Single-Cell mRNA-Seq of In Vitro-Derived Human Neurons Using Smart-Seq2

Christoph Schweingruber; Jik Nijssen; Julio Aguila Benitez; Eva Hedlund

doi:10.1007/978-1-0716-2815-7_11

Single-Cell mRNA-Seq of In Vitro-Derived Human Neurons Using Smart-Seq2

Methods Mol Biol. 2023:2594:143-164. doi: 10.1007/978-1-0716-2815-7_11.

Authors

Christoph Schweingruber^{1

2}, Jik Nijssen², Julio Aguila Benitez³, Eva Hedlund^{4

5}

Affiliations

¹ Department for Biochemistry and Biophysics, Stockholm University, Stockholm, Sweden.
² Department of Cell and Molecular Biology, Karolinska Institutet, Stockholm, Sweden.
³ Department of Neuroscience, Karolinska Institutet, Stockholm, Sweden.
⁴ Department for Biochemistry and Biophysics, Stockholm University, Stockholm, Sweden. [email protected].
⁵ Department of Cell and Molecular Biology, Karolinska Institutet, Stockholm, Sweden. [email protected].

PMID: 36264494
DOI: 10.1007/978-1-0716-2815-7_11

Abstract

Single-cell mRNA sequencing can dissect heterogeneous cell populations as it can identify cell types and cellular states based on their unique transcriptional signatures. We use fluorescence-activated cell sorting (FACS) to isolate individual cultured neurons derived from human-induced pluripotent stem cells (hiPSCs) followed by polyA-based Smart-Seq2 RNA sequencing to analyze the single-cell transcriptional profiles. We provide protocols and guidelines on dissociation, cell selection, and library preparation that can be readily adapted to other cell types or tissue samples.

Keywords: FACS; Motor neuron; Single-cell RNA sequencing; Smart-Seq2.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Gene Expression Profiling / methods
Gene Library
Humans
Neurons* / metabolism
RNA, Messenger / genetics
RNA, Messenger / metabolism
Sequence Analysis, RNA / methods
Single-Cell Analysis* / methods

Substances

RNA, Messenger