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Isolation, culture, and immunostaining of neonatal rat ventricular myocytes

2021; Elsevier BV; Volume: 2; Issue: 4 Linguagem: Inglês

10.1016/j.xpro.2021.100950

2666-1667

Ana Helena Macedo Pereira, Alisson C. Cardoso, Kleber G. Franchini,

Mechanical Circulatory Support Devices

Isolation and culture of ventricular cardiomyocytes from neonatal rats (NRVMs) is a powerful model to study neonatal cardiac development, cell cycle regulation, and cardiac physiology and pathology in vitro. Here, we present our modified enzymatic digestion protocol followed by two-step discontinuous Percoll gradient centrifugation to isolate a high yield of viable ventricular cardiomyocytes from neonatal rats. Finally, here we describe an immunostaining protocol for cytosolic and nuclear staining of NRVMs. For complete details on the use and execution of this protocol, please refer to Pereira et al. (2020).