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

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

STAR Protocols (Dec 2021)

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

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

Affiliations

Gabriella E. Farrugia: Baker Heart and Diabetes Institute, Melbourne, VIC 3004, Australia
Micheal A. McLellan: The Jackson Laboratory, Bar Harbor, ME, USA; Tufts University, Graduate School of Biomedical Sciences, Boston, MA, USA
Kate L. Weeks: Baker 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, Australia
Aya Matsumoto: Baker Heart and Diabetes Institute, Melbourne, VIC 3004, Australia
Charles D. Cohen: Baker Heart and Diabetes Institute, Melbourne, VIC 3004, Australia; Centre for Cardiovascular Biology and Disease Research, La Trobe University, Melbourne, VIC 3083, Australia
Crisdion Krstevski: Baker Heart and Diabetes Institute, Melbourne, VIC 3004, Australia; Centre for Cardiovascular Biology and Disease Research, La Trobe University, Melbourne, VIC 3083, Australia
Taylah L. Gaynor: Baker Heart and Diabetes Institute, Melbourne, VIC 3004, Australia; Centre for Cardiovascular Biology and Disease Research, La Trobe University, Melbourne, VIC 3083, Australia
Adam C. Parslow: Baker Heart and Diabetes Institute, Melbourne, VIC 3004, Australia; Baker Department of Cardiometabolic Health, The University of Melbourne, Parkville, VIC 3010, Australia
Julie R. McMullen: Baker 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, Australia
Alexander R. Pinto: Baker 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 author

Journal volume & issue: Vol. 2, no. 4
p. 100866

Abstract

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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).

Published in STAR Protocols

ISSN: 2666-1667 (Online)
Publisher: Elsevier
Country of publisher: United States
LCC subjects: Science: Science (General)
Website: https://star-protocols.cell.com/

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Abstract

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