Cargando…
A protocol for rapid and parallel isolation of myocytes and non-myocytes from multiple mouse hearts
This protocol features parallel isolation of myocytes and non-myocytes from murine hearts. It was designed with considerations for (1) time required to extract cardiac cells, (2) cell viability, and (3) protocol scalability. Here, a peristaltic pump and 3D-printed elements are combined to perfuse th...
Autores principales: | , , , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier
2021
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8496295/ https://www.ncbi.nlm.nih.gov/pubmed/34647038 http://dx.doi.org/10.1016/j.xpro.2021.100866 |
_version_ | 1784579726873985024 |
---|---|
author | Farrugia, Gabriella E. McLellan, Micheal A. Weeks, Kate L. Matsumoto, Aya Cohen, Charles D. Krstevski, Crisdion Gaynor, Taylah L. Parslow, Adam C. McMullen, Julie R. Pinto, Alexander R. |
author_facet | Farrugia, Gabriella E. McLellan, Micheal A. Weeks, Kate L. Matsumoto, Aya Cohen, Charles D. Krstevski, Crisdion Gaynor, Taylah L. Parslow, Adam C. McMullen, Julie R. Pinto, Alexander R. |
author_sort | Farrugia, Gabriella E. |
collection | PubMed |
description | This protocol features parallel isolation of myocytes and non-myocytes from murine hearts. It was designed with considerations for (1) time required to extract cardiac cells, (2) cell viability, and (3) protocol scalability. Here, a peristaltic pump and 3D-printed elements are combined to perfuse the heart with enzymes to dissociate cells. Myocytes and non-myocytes extracted using this protocol are separated by centrifugation and/or fluorescence-activated cell sorting for use in downstream applications including single-cell omics or other bio-molecular analyses. For complete details on the use and execution of this protocol, please refer to McLellan et al. (2020). |
format | Online Article Text |
id | pubmed-8496295 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
spelling | pubmed-84962952021-10-12 A protocol for rapid and parallel isolation of myocytes and non-myocytes from multiple mouse hearts Farrugia, Gabriella E. McLellan, Micheal A. Weeks, Kate L. Matsumoto, Aya Cohen, Charles D. Krstevski, Crisdion Gaynor, Taylah L. Parslow, Adam C. McMullen, Julie R. Pinto, Alexander R. STAR Protoc Protocol This protocol features parallel isolation of myocytes and non-myocytes from murine hearts. It was designed with considerations for (1) time required to extract cardiac cells, (2) cell viability, and (3) protocol scalability. Here, a peristaltic pump and 3D-printed elements are combined to perfuse the heart with enzymes to dissociate cells. Myocytes and non-myocytes extracted using this protocol are separated by centrifugation and/or fluorescence-activated cell sorting for use in downstream applications including single-cell omics or other bio-molecular analyses. For complete details on the use and execution of this protocol, please refer to McLellan et al. (2020). Elsevier 2021-09-30 /pmc/articles/PMC8496295/ /pubmed/34647038 http://dx.doi.org/10.1016/j.xpro.2021.100866 Text en © 2021 The Author(s) https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). |
spellingShingle | Protocol Farrugia, Gabriella E. McLellan, Micheal A. Weeks, Kate L. Matsumoto, Aya Cohen, Charles D. Krstevski, Crisdion Gaynor, Taylah L. Parslow, Adam C. McMullen, Julie R. Pinto, Alexander R. A protocol for rapid and parallel isolation of myocytes and non-myocytes from multiple mouse hearts |
title | A protocol for rapid and parallel isolation of myocytes and non-myocytes from multiple mouse hearts |
title_full | A protocol for rapid and parallel isolation of myocytes and non-myocytes from multiple mouse hearts |
title_fullStr | A protocol for rapid and parallel isolation of myocytes and non-myocytes from multiple mouse hearts |
title_full_unstemmed | A protocol for rapid and parallel isolation of myocytes and non-myocytes from multiple mouse hearts |
title_short | A protocol for rapid and parallel isolation of myocytes and non-myocytes from multiple mouse hearts |
title_sort | protocol for rapid and parallel isolation of myocytes and non-myocytes from multiple mouse hearts |
topic | Protocol |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8496295/ https://www.ncbi.nlm.nih.gov/pubmed/34647038 http://dx.doi.org/10.1016/j.xpro.2021.100866 |
work_keys_str_mv | AT farrugiagabriellae aprotocolforrapidandparallelisolationofmyocytesandnonmyocytesfrommultiplemousehearts AT mclellanmicheala aprotocolforrapidandparallelisolationofmyocytesandnonmyocytesfrommultiplemousehearts AT weekskatel aprotocolforrapidandparallelisolationofmyocytesandnonmyocytesfrommultiplemousehearts AT matsumotoaya aprotocolforrapidandparallelisolationofmyocytesandnonmyocytesfrommultiplemousehearts AT cohencharlesd aprotocolforrapidandparallelisolationofmyocytesandnonmyocytesfrommultiplemousehearts AT krstevskicrisdion aprotocolforrapidandparallelisolationofmyocytesandnonmyocytesfrommultiplemousehearts AT gaynortaylahl aprotocolforrapidandparallelisolationofmyocytesandnonmyocytesfrommultiplemousehearts AT parslowadamc aprotocolforrapidandparallelisolationofmyocytesandnonmyocytesfrommultiplemousehearts AT mcmullenjulier aprotocolforrapidandparallelisolationofmyocytesandnonmyocytesfrommultiplemousehearts AT pintoalexanderr aprotocolforrapidandparallelisolationofmyocytesandnonmyocytesfrommultiplemousehearts AT farrugiagabriellae protocolforrapidandparallelisolationofmyocytesandnonmyocytesfrommultiplemousehearts AT mclellanmicheala protocolforrapidandparallelisolationofmyocytesandnonmyocytesfrommultiplemousehearts AT weekskatel protocolforrapidandparallelisolationofmyocytesandnonmyocytesfrommultiplemousehearts AT matsumotoaya protocolforrapidandparallelisolationofmyocytesandnonmyocytesfrommultiplemousehearts AT cohencharlesd protocolforrapidandparallelisolationofmyocytesandnonmyocytesfrommultiplemousehearts AT krstevskicrisdion protocolforrapidandparallelisolationofmyocytesandnonmyocytesfrommultiplemousehearts AT gaynortaylahl protocolforrapidandparallelisolationofmyocytesandnonmyocytesfrommultiplemousehearts AT parslowadamc protocolforrapidandparallelisolationofmyocytesandnonmyocytesfrommultiplemousehearts AT mcmullenjulier protocolforrapidandparallelisolationofmyocytesandnonmyocytesfrommultiplemousehearts AT pintoalexanderr protocolforrapidandparallelisolationofmyocytesandnonmyocytesfrommultiplemousehearts |