A protocol for rapid and parallel isolation of myocytes and non-myocytes from multiple mouse hearts

A protocol for rapid and parallel isolation of myocytes and non-myocytes from multiple mouse hearts

Summary: 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 p...

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Main Authors: Gabriella E. Farrugia, Micheal A. McLellan, Kate L. Weeks, Aya Matsumoto, Charles D. Cohen, Crisdion Krstevski, Taylah L. Gaynor, Adam C. Parslow, Julie R. McMullen, Alexander R. Pinto
Format: Article
Language:English
Published: Elsevier 2021-12-01
Series:STAR Protocols
Subjects:
Cell Biology
Cell isolation
Cell separation/fractionation
Online Access:http://www.sciencedirect.com/science/article/pii/S2666166721005724
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spelling doaj-e554ac6acc834cce8edb2df50ff09a7e2021-10-03T04:44:04ZengElsevierSTAR Protocols2666-16672021-12-0124100866A protocol for rapid and parallel isolation of myocytes and non-myocytes from multiple mouse heartsGabriella E. Farrugia0Micheal A. McLellan1Kate L. Weeks2Aya Matsumoto3Charles D. Cohen4Crisdion Krstevski5Taylah L. Gaynor6Adam C. Parslow7Julie R. McMullen8Alexander R. Pinto9Baker Heart and Diabetes Institute, Melbourne, VIC 3004, AustraliaThe Jackson Laboratory, Bar Harbor, ME, USA; Tufts University, Graduate School of Biomedical Sciences, Boston, MA, USABaker Heart and Diabetes Institute, Melbourne, VIC 3004, Australia; Department of Diabetes, Central Clinical School, Monash University, Clayton, VIC 3800, Australia; Baker Department of Cardiometabolic Health, The University of Melbourne, Parkville, VIC 3010, AustraliaBaker Heart and Diabetes Institute, Melbourne, VIC 3004, AustraliaBaker Heart and Diabetes Institute, Melbourne, VIC 3004, Australia; Centre for Cardiovascular Biology and Disease Research, La Trobe University, Melbourne, VIC 3083, AustraliaBaker Heart and Diabetes Institute, Melbourne, VIC 3004, Australia; Centre for Cardiovascular Biology and Disease Research, La Trobe University, Melbourne, VIC 3083, AustraliaBaker Heart and Diabetes Institute, Melbourne, VIC 3004, Australia; Centre for Cardiovascular Biology and Disease Research, La Trobe University, Melbourne, VIC 3083, AustraliaBaker Heart and Diabetes Institute, Melbourne, VIC 3004, Australia; Baker Department of Cardiometabolic Health, The University of Melbourne, Parkville, VIC 3010, AustraliaBaker Heart and Diabetes Institute, Melbourne, VIC 3004, Australia; Department of Diabetes, Central Clinical School, Monash University, Clayton, VIC 3800, Australia; Centre for Cardiovascular Biology and Disease Research, La Trobe University, Melbourne, VIC 3083, Australia; Department of Physiology and Department of Medicine Alfred Hospital, Monash University, Melbourne, VIC, Australia; Department of Physiology, Anatomy and Microbiology, La Trobe University, Bundoora, VIC, AustraliaBaker Heart and Diabetes Institute, Melbourne, VIC 3004, Australia; Centre for Cardiovascular Biology and Disease Research, La Trobe University, Melbourne, VIC 3083, Australia; Department of Physiology, Anatomy and Microbiology, La Trobe University, Bundoora, VIC, Australia; Corresponding authorSummary: 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).http://www.sciencedirect.com/science/article/pii/S2666166721005724Cell BiologyCell isolationCell separation/fractionation
collection DOAJ
language English
format Article
sources DOAJ
author Gabriella E. Farrugia
Micheal A. McLellan
Kate L. Weeks
Aya Matsumoto
Charles D. Cohen
Crisdion Krstevski
Taylah L. Gaynor
Adam C. Parslow
Julie R. McMullen
Alexander R. Pinto
spellingShingle Gabriella E. Farrugia
Micheal A. McLellan
Kate L. Weeks
Aya Matsumoto
Charles D. Cohen
Crisdion Krstevski
Taylah L. Gaynor
Adam C. Parslow
Julie R. McMullen
Alexander R. Pinto
A protocol for rapid and parallel isolation of myocytes and non-myocytes from multiple mouse hearts
STAR Protocols
Cell Biology
Cell isolation
Cell separation/fractionation
author_facet Gabriella E. Farrugia
Micheal A. McLellan
Kate L. Weeks
Aya Matsumoto
Charles D. Cohen
Crisdion Krstevski
Taylah L. Gaynor
Adam C. Parslow
Julie R. McMullen
Alexander R. Pinto
author_sort Gabriella E. Farrugia
title 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_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_sort protocol for rapid and parallel isolation of myocytes and non-myocytes from multiple mouse hearts
publisher Elsevier
series STAR Protocols
issn 2666-1667
publishDate 2021-12-01
description Summary: 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).
topic Cell Biology
Cell isolation
Cell separation/fractionation
url http://www.sciencedirect.com/science/article/pii/S2666166721005724
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