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...

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Detalles Bibliográficos
Autores principales: Santoro, Federica, Chien, Kenneth R., Sahara, Makoto
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2021
Materias:
Protocol
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
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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).
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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
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AT saharamakoto isolationofhumanescderivedcardiacderivativesandembryonicheartcellsforpopulationandsinglecellrnaseqanalysis

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