Isolation and Culture of Adult Mouse Cardiomyocytes for Cell Signaling and <em>in vitro</em> Cardiac Hypertrophy

Abstract

Technological advances have made genetically modified mice, including transgenic and gene knockout mice, an essential tool in many research fields. Adult cardiomyocytes are widely accepted as a good model for cardiac cellular physiology and pathophysiology, as well as for pharmaceutical intervention. Genetically modified mice preclude the need for complicated cardiomyocyte infection processes to generate the desired genotype, which are inefficient due to cardiomyocytes' terminal differentiation. Isolation and culture of high quantity and quality functional cardiomyocytes will dramatically benefit cardiovascular research and provide an important tool for cell signaling transduction research and drug development. Here, we describe a well-established method for isolation of adult mouse cardiomyocytes that can be implemented with little training. The mouse heart is excised and cannulated to an isolated heart system, then perfused with a calcium-free and high potassium buffer followed by type II collagenase digestion in Langendorff retrograde perfusion mode. This protocol yields a consistent result for the collection of functional adult mouse cardiomyocytes from a variety of genetically modified mice.

Targeting MRTF/SRF in CAP2-dependent dilated cardiomyopathy delays disease onset

“…Cardiomyocyte isolation. Cardiomyocytes were isolated by the Langendorff method, using retrograde perfusion through the aorta with enzyme-containing solutions, as described (51). Briefly, mice were anesthetized with an i.p.…”

Mesenchymal Stem Cells Promote Cardiac Muscle Repair via Enhanced Neovascularization

“…The mouse hearts were digested into single cells as has been described before [19]. The digests were incubated either with M-cadherin antibody (M-Cad, Millipore, Billerica, MA, USA), or with CD31 antibody (Becton-Dickinson Biosciences), or F4/80 (ebioscience, San Diego, CA, USA) followed by labeling with PE-conjugated anti-mouse second antibody (Becton-Dickinson Biosciences).…”

MITF interacts with the SWI/SNF subunit, BRG1, to promote GATA4 expression in cardiac hypertrophy

“…Cardiomyocytes were isolated from hearts obtained from 3 month old male mice as described in [52]. 10μM isoproterenol or 20nM angiotensin II was used to stimulate hypertrophy.…”