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

doi:10.1016/j.xpro.2021.100866

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

STAR Protoc. 2021 Sep 30;2(4):100866. doi: 10.1016/j.xpro.2021.100866. eCollection 2021 Dec 17.

Authors

Gabriella E Farrugia¹, Micheal A McLellan^{2

3}, Kate L Weeks^{1

4

5}, Aya Matsumoto¹, Charles D Cohen^{1

6}, Crisdion Krstevski^{1

6}, Taylah L Gaynor^{1

6}, Adam C Parslow^{1

5}, Julie R McMullen^{1

4

6

7

8}, Alexander R Pinto^{1

6

8}

Affiliations

¹ Baker Heart and Diabetes Institute, Melbourne, VIC 3004, Australia.
² The Jackson Laboratory, Bar Harbor, ME, USA.
³ Tufts University, Graduate School of Biomedical Sciences, Boston, MA, USA.
⁴ Department of Diabetes, Central Clinical School, Monash University, Clayton, VIC 3800, Australia.
⁵ Baker Department of Cardiometabolic Health, The University of Melbourne, Parkville, VIC 3010, Australia.
⁶ Centre for Cardiovascular Biology and Disease Research, La Trobe University, Melbourne, VIC 3083, Australia.
⁷ Department of Physiology and Department of Medicine Alfred Hospital, Monash University, Melbourne, VIC, Australia.
⁸ Department of Physiology, Anatomy and Microbiology, La Trobe University, Bundoora, VIC, Australia.

Abstract

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).

Keywords: Cell Biology; Cell isolation; Cell separation/fractionation.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Cell Culture Techniques
Cell Separation / methods*
Cells, Cultured
Genomics
Male
Mice
Mice, Inbred C57BL
Myocardium / cytology*
Myocytes, Cardiac* / classification
Myocytes, Cardiac* / cytology
Single-Cell Analysis / methods*