Isolation of human ESC-derived cardiac derivatives and embryonic heart cells for population and single-cell RNA-seq analysis

STAR Protocols

Makoto Sahara¹  Kenneth R. Chien²  Federica Santoro² 

[1] Department of Medicine, Karolinska Institutet, Stockholm 141 86, Sweden;Department of Cell and Molecular Biology, Karolinska Institutet, Stockholm 171 77, Sweden;

关键词: Cell differentiation;    Cell isolation;    Flow cytometry/mass cytometry;    RNA-seq;    Single cell;    Stem cells;   

DOI  :  

来源: DOAJ

【 摘 要 】

Summary: The combination of population and single-cell RNA sequencing analysis using human embryonic stem cell (hESC) differentiation and developmental tissues is a powerful approach to elucidate an organ-specific cellular and molecular atlas in human embryogenesis. This protocol describes (1) cardiac-directed differentiation and isolation of hESC-derived cardiac derivatives with fluorescence-activated cell sorting, (2) isolation of human embryonic heart-derived single cardiac cells, and (3) construction of cDNA libraries with Smart-seq2. These allow for the preparation of human developmental samples for comprehensive transcriptional analysis.For complete details on the use and execution of this protocol, please refer to Sahara et al. (2019).

【 授权许可】

Unknown