Isolation of Adult Mouse Cardiomyocytes Using Langendorff Perfusion Apparatus

Abstract

Heart disease is one of the leading causes of death in the United States. Isolation and culture adult cardiomyocytes are important for studying cardiomyocyte contractility, heart hypertrophy, and cardiac failure. In contrast to neonatal cardiomyocyte isolation, adult mice cardiomyocytes isolation is challenging due to firm connections among cardiomyocytes through intercalated discs. The availability of newly generated genetically modified mouse lines requires to establish protocols to isolation and culture adult mouse cardiomyocyte for in vitro studies. In this manuscript, we described a straightforward method of isolating adult mouse cardiomyocytes using Langendorff perfusion apparatus. Briefly, the hearts were harvested from adult mice and the heart was mounted to Lagendorff apparatus. After perfusion with calcium depletion and collagenase digestion, the left ventricles were minced and filtered. Lastly, the separated cardiomyocytes were treated with CaCl₂. The isolated cardiac myocytes can be utilized in a broad range of experiments including screening for drugs.

Keywords: Adult cardiomyocytes isolation; Langendorff; Trypsin.

Fig. 1

Langendorff perfusion system

Fig. 2

Surgical tools for isolating adult heart and loading heart into Langendorff apparatus

Fig. 3

Heart cannulation system

Fig. 4

50 ml Falcon tube with Sefar Nylon Mesh filter top for accumulating filtered cardiomyocytes

Fig. 5

Heart cannulation

Fig. 6

Isolated adult cardiomyocytes