Isolation, Culture and Transduction of Adult Mouse Cardiomyocytes

Abstract

Cultured cardiomyocytes can be used to study cardiomyocyte biology using techniques that are complementary to in vivo systems. For example, the purity and accessibility of in vitro culture enables fine control over biochemical analyses, live imaging, and electrophysiology. Long-term culture of cardiomyocytes offers access to additional experimental approaches that cannot be completed in short term cultures. For example, the in vitro investigation of dedifferentiation, cell cycle re-entry, and cell division has thus far largely been restricted to rat cardiomyocytes, which appear to be more robust in long-term culture. However, the availability of a rich toolset of transgenic mouse lines and well-developed disease models make mouse systems attractive for cardiac research. Although several reports exist of adult mouse cardiomyocyte isolation, few studies demonstrate their long-term culture. Presented here, is a step-by-step method for the isolation and long-term culture of adult mouse cardiomyocytes. First, retrograde Langendorff perfusion is used to efficiently digest the heart with proteases, followed by gravity sedimentation purification. After a period of dedifferentiation following isolation, the cells gradually attach to the culture and can be cultured for weeks. Adenovirus cell lysate is used to efficiently transduce the isolated cardiomyocytes. These methods provide a simple, yet powerful model system to study cardiac biology.

Direct Comparison of Mononucleated and Binucleated Cardiomyocytes Reveals Molecular Mechanisms Underlying Distinct Proliferative Competencies

“…Isolation of Cardiomyocytes for FACS Isolation of Adult Cardiomyocytes Cardiomyocytes were isolated from mice aged 8-13 weeks using a protocol modified from methods previously described (Judd et al, 2016;Robison et al, 2016;Tian et al, 2015). Buffers were prepared on the day of isolation as follows.…”

Multinucleated polyploid cardiomyocytes undergo an enhanced adaptability to hypoxia via mitophagy

“…Adult cardiomyocytes (CMs) in the left ventricular wall obtained from 3 months old male mice were isolated as previously reported [8]. Ventricles (three hearts per sample) were harvested from hypoxic or normoxic mice for 7 and 28 days and perfused with a cardiomyocyte isolation buffer using a Langendorff system for~3 min.…”

Novel microfluidic platform accelerates processing of tissue into single cells for molecular analysis and primary culture models

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