Cargando…
Isolation of human ESC-derived cardiac derivatives and embryonic heart cells for population and single-cell RNA-seq analysis
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-direct...
Autores principales: | , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier
2021
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7887647/ https://www.ncbi.nlm.nih.gov/pubmed/33644774 http://dx.doi.org/10.1016/j.xpro.2021.100339 |
_version_ | 1783652025869795328 |
---|---|
author | Santoro, Federica Chien, Kenneth R. Sahara, Makoto |
author_facet | Santoro, Federica Chien, Kenneth R. Sahara, Makoto |
author_sort | Santoro, Federica |
collection | PubMed |
description | 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). |
format | Online Article Text |
id | pubmed-7887647 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
spelling | pubmed-78876472021-02-26 Isolation of human ESC-derived cardiac derivatives and embryonic heart cells for population and single-cell RNA-seq analysis Santoro, Federica Chien, Kenneth R. Sahara, Makoto STAR Protoc Protocol 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). Elsevier 2021-02-11 /pmc/articles/PMC7887647/ /pubmed/33644774 http://dx.doi.org/10.1016/j.xpro.2021.100339 Text en © 2021 The Author(s) http://creativecommons.org/licenses/by/4.0/ This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Protocol Santoro, Federica Chien, Kenneth R. Sahara, Makoto Isolation of human ESC-derived cardiac derivatives and embryonic heart cells for population and single-cell RNA-seq analysis |
title | Isolation of human ESC-derived cardiac derivatives and embryonic heart cells for population and single-cell RNA-seq analysis |
title_full | Isolation of human ESC-derived cardiac derivatives and embryonic heart cells for population and single-cell RNA-seq analysis |
title_fullStr | Isolation of human ESC-derived cardiac derivatives and embryonic heart cells for population and single-cell RNA-seq analysis |
title_full_unstemmed | Isolation of human ESC-derived cardiac derivatives and embryonic heart cells for population and single-cell RNA-seq analysis |
title_short | Isolation of human ESC-derived cardiac derivatives and embryonic heart cells for population and single-cell RNA-seq analysis |
title_sort | isolation of human esc-derived cardiac derivatives and embryonic heart cells for population and single-cell rna-seq analysis |
topic | Protocol |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7887647/ https://www.ncbi.nlm.nih.gov/pubmed/33644774 http://dx.doi.org/10.1016/j.xpro.2021.100339 |
work_keys_str_mv | AT santorofederica isolationofhumanescderivedcardiacderivativesandembryonicheartcellsforpopulationandsinglecellrnaseqanalysis AT chienkennethr isolationofhumanescderivedcardiacderivativesandembryonicheartcellsforpopulationandsinglecellrnaseqanalysis AT saharamakoto isolationofhumanescderivedcardiacderivativesandembryonicheartcellsforpopulationandsinglecellrnaseqanalysis |