利用 Smart-Seq2 对体外诱导的人类神经元进行单细胞 mRNA-Seq 测序。

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

机构信息

Department for Biochemistry and Biophysics, Stockholm University, Stockholm, Sweden.

Department of Cell and Molecular Biology, Karolinska Institutet, Stockholm, Sweden.

出版信息

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

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

PMID:36264494

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.

摘要

单细胞 mRNA 测序可以对异质细胞群体进行剖析，因为它可以根据细胞的独特转录特征来鉴定细胞类型和细胞状态。我们使用荧光激活细胞分选（FACS）分离来自人诱导多能干细胞（hiPSC）的单个培养神经元，然后进行基于 polyA 的 Smart-Seq2 RNA 测序，以分析单细胞转录谱。我们提供了关于解离、细胞选择和文库制备的方案和指南，这些方案和指南可以很容易地适应其他细胞类型或组织样本。