Single-Cardiomyocyte RNA Sequencing to Dissect the Molecular Pathophysiology of the Heart

Methods Mol Biol. 2021;2320:183-192. doi: 10.1007/978-1-0716-1484-6_18.


Heart failure is caused by a complicated pathogenic process and has a poor prognosis. Quality of life is often impaired due to repeated hospitalization. Integrative analysis of the morphological, physiological, and molecular profiles of cardiomyocytes, which are responsible mainly for heart contraction, may lead to a deeper understanding of the pathogenesis of heart failure. However, unlike other types of cells, cardiomyocytes are relatively large, vulnerable to stress, and difficult to use for single-cell analysis. With some ingenuity, we have established a single-cardiomyocyte analysis pipeline. Here, we describe the procedure for single-cell RNA sequencing of adult mouse cardiomyocytes from isolation to analysis.

Keywords: Cardiomyocytes; Full-length cDNA library construction; Langendorff perfusion; Single-cell RNA-seq.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Equipment Design
  • Gene Library
  • Heart / physiopathology*
  • Heart Failure / physiopathology
  • Male
  • Mice
  • Myocardium / chemistry*
  • Myocytes, Cardiac / chemistry*
  • Perfusion / instrumentation
  • Perfusion / methods
  • RNA-Seq / methods*
  • Reverse Transcriptase Polymerase Chain Reaction / methods
  • Single-Cell Analysis / methods*
  • Specimen Handling