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Isolation, culture, and immunostaining of neonatal rat ventricular myocytes.


ABSTRACT: 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).

SUBMITTER: Pereira AHM 

PROVIDER: S-EPMC8599495 | biostudies-literature | 2021 Dec

REPOSITORIES: biostudies-literature

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Isolation, culture, and immunostaining of neonatal rat ventricular myocytes.

Pereira Ana Helena Macedo AHM   Cardoso Alisson Campos AC   Franchini Kleber Gomes KG  

STAR protocols 20211112 4


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 <i>in vitro</i>. 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  ...[more]

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