Simple Method to Isolate Individual Mouse Cardiomyocytes Using Enzymatic Digestion via Retrograde Perfusion

Madyson O Hintz; Ute Zellhuber-McMillan; Peter P Jones

doi:10.1007/978-1-0716-4342-6_2

Simple Method to Isolate Individual Mouse Cardiomyocytes Using Enzymatic Digestion via Retrograde Perfusion

Methods Mol Biol. 2025:2894:7-20. doi: 10.1007/978-1-0716-4342-6_2.

Authors

Madyson O Hintz¹, Ute Zellhuber-McMillan¹, Peter P Jones²

Affiliations

¹ Department of Physiology and HeartOtago, School of Biomedical Sciences, University of Otago, Otago, New Zealand.
² Department of Physiology and HeartOtago, School of Biomedical Sciences, University of Otago, Otago, New Zealand. [email protected].

PMID: 39699806
DOI: 10.1007/978-1-0716-4342-6_2

Abstract

Isolated single cardiomyocytes are commonly used to understand fundamental cardiac cell physiology and cell signaling at the molecular level. The protocol to isolate cardiomyocytes is under continual optimization between laboratories, despite being conducted for many years. The delicate nature of these cells coinciding with laboratory equipment variations can make the consistency of isolation challenging. The protocol described here presents a simple method to routinely isolate high-quality cardiomyocytes using Langendorff perfusion. This method provides cells well suited for analysis of physiological cellular calcium handling and spontaneous calcium release.

Keywords: Cardiac physiologyCardiac physiology; Cardiomyocytes; Cell isolation; Langendorff.

MeSH terms

Animals
Calcium / metabolism
Cell Separation* / methods
Mice
Myocytes, Cardiac* / cytology
Myocytes, Cardiac* / metabolism
Perfusion* / instrumentation
Perfusion* / methods

Substances

Calcium