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

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

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) cardi...

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Bibliographic Details
Main Authors: Federica Santoro, Kenneth R. Chien, Makoto Sahara
Format: Article
Language:English
Published: Elsevier 2021-03-01
Series:STAR Protocols
Subjects:
Cell differentiation
Cell isolation
Flow cytometry/mass cytometry
RNA-seq
Single cell
Stem cells
Online Access:http://www.sciencedirect.com/science/article/pii/S2666166721000460
Description
Summary: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).
ISSN:2666-1667

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