Protocol for simultaneous isolation of high-quality and high-quantity cardiomyocytes and non-myocyte cells from adult rat hearts

• Published in: STAR protocols Vol. 5; no. 3; p. 103174
• Main Authors: Zimmer, Alexsandra, Wang, Eric R., Choudhary, Gaurav, Zhang, Peng
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 20-09-2024; Elsevier
• Subjects: Animals; Cell Biology; Cell culture; Cell isolation; Cell Separation - methods; Cell separation/fractionation; Cell-based Assays; Flow Cytometry; Flow Cytometry - methods; Myocardium - cytology; Myocytes, Cardiac - cytology; Protocol; Rats; Single Cell; Cell culture; Flow Cytometry; Cell-based Assays; Cell separation/fractionation; Single Cell; Cell isolation; Cell Biology
• ISSN: 2666-1667; 2666-1667
• DOI: 10.1016/j.xpro.2024.103174

Isolating high-quality different cell types is a powerful approach for understanding cellular compositions and features in the heart, but it is challenging. The available protocols typically focus on isolating one or two cell types. Here, we present a protocol to simultaneously isolate high-quality and high-quantity cardiomyocytes and non-myocyte cells, including immune cells, from adult rat hearts. We describe steps for purifying cells using bovine serum albumin. We also detail procedures for viability analysis and cell type identification using fluorescence-activated cell sorting.
For complete details on the use and execution of this protocol, please refer to Zhang et al.,1 Valkov et al.,2 Vang et al.,3 and Li et al.4
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•Protocol to isolate cardiac cells using Langendorff perfusion and enzymatic digestion•Steps to purify cardiomyocytes and non-myocyte cells using bovine serum albumin•Steps for medium preparation and culturing high-quality cardiomyocytes and fibroblasts•Characterization of non-myocyte cells using fluorescence-activated cell sorting
Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.
Isolating high-quality different cell types is a powerful approach for understanding cellular compositions and features in the heart, but it is challenging. The available protocols typically focus on isolating one or two cell types. Here, we present a protocol to simultaneously isolate high-quality and high-quantity cardiomyocytes and non-myocyte cells, including immune cells, from adult rat hearts. We describe steps for purifying cells using bovine serum albumin. We also detail procedures for viability analysis and cell type identification using fluorescence-activated cell sorting.