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.

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