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用于群体和单细胞 RNA-seq 分析的人 ESC 来源的心脏衍生物和胚胎心脏细胞的分离。

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

机构信息

Department of Cell and Molecular Biology, Karolinska Institutet, Stockholm 171 77, Sweden.

Department of Medicine, Karolinska Institutet, Stockholm 141 86, Sweden.

出版信息

STAR Protoc. 2021 Feb 11;2(1):100339. doi: 10.1016/j.xpro.2021.100339. eCollection 2021 Mar 19.

DOI:10.1016/j.xpro.2021.100339
PMID:33644774
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7887647/
Abstract

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).

摘要

使用人类胚胎干细胞 (hESC) 分化和发育组织进行人群和单细胞 RNA 测序分析是阐明人类胚胎发生中特定器官的细胞和分子图谱的有力方法。本方案描述了 (1) 心脏定向分化和荧光激活细胞分选分离 hESC 衍生的心脏衍生物，(2) 分离人类胚胎心脏来源的单个心脏细胞，以及 (3) 使用 Smart-seq2 构建 cDNA 文库。这些方法为全面转录分析准备了人类发育样本。有关该方案使用和执行的完整详细信息，请参阅 Sahara 等人。(2019)。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/aa93/7887647/a57b6a60c318/gr5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/aa93/7887647/22ff32e1c61e/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/aa93/7887647/8f8855e61782/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/aa93/7887647/fc3a914303e1/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/aa93/7887647/6a624a5e5929/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/aa93/7887647/b31214147e33/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/aa93/7887647/a57b6a60c318/gr5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/aa93/7887647/22ff32e1c61e/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/aa93/7887647/8f8855e61782/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/aa93/7887647/fc3a914303e1/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/aa93/7887647/6a624a5e5929/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/aa93/7887647/b31214147e33/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/aa93/7887647/a57b6a60c318/gr5.jpg

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