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.


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*