Key Laboratory of Cardiovascular Intervention and Regenerative Medicine of Zhejiang Province 1 article published in JoVE Medicine Preparation of a Non-Cardiomyocyte Cell Suspension for Single-Cell RNA Sequencing from a Post-Myocardial Infarction Adult Mouse Heart Chengchen Huang1,2, Jiahe Han1,2, Shuo Sun1,2, Guosheng Fu*1,2, Min Shang*1,2 1Key Laboratory of Cardiovascular Intervention and Regenerative Medicine of Zhejiang Province, 2Department of Cardiology, Sir Run Run Shaw Hospital, School of Medicine, Zhejiang University Hangzhou Here, we describe a protocol to isolate a sufficient amount of single non-cardiomyocytes with high viability from a post-myocardial infarction (MI) mouse heart. This can be used for subsequent single-cell sequencing, flow cytometry analysis, and primary cell culture.