Journal article
A protocol for rapid and parallel isolation of myocytes and non-myocytes from multiple mouse hearts
GE Farrugia, MA McLellan, KL Weeks, A Matsumoto, CD Cohen, C Krstevski, TL Gaynor, AC Parslow, JR McMullen, AR Pinto
STAR Protocols | Published : 2021
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).
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Grants
Awarded by State Government of Victoria
Funding Acknowledgements
This work is supported by the National Health and Medical Research Council (Australia) Ideas Grant (GNT1188503) to A.R.P. and in part by the Victorian Government's Operational Infrastructure Support Program. K.L.W. is supported by a Future Leader Fellowship from the National Heart Foundation of Australia (award ID 102539) . J.R.M. is supported by a National Health and Medical Research Council Senior Research Fellowship (ID 1078985) . The authors acknowledge the facilities and the scientific and technical assistance of the Baker Heart and Diabetes Institute Microscopy Platform.