Isolation, culture, and immunostaining of neonatal rat ventricular myocytes

Ana Helena Macedo Pereira; Alisson Campos Cardoso; Kleber Gomes Franchini

doi:10.1016/j.xpro.2021.100950

Isolation, culture, and immunostaining of neonatal rat ventricular myocytes

STAR Protoc. 2021 Nov 12;2(4):100950. doi: 10.1016/j.xpro.2021.100950. eCollection 2021 Dec 17.

Authors

Ana Helena Macedo Pereira¹, Alisson Campos Cardoso¹, Kleber Gomes Franchini¹

Affiliation

¹ Brazilian Biosciences National Laboratory, Brazilian Center for Research in Energy and Materials (CNPEM), Campinas, Sao Paulo, Brazil.

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).

Keywords: Cell Biology; Cell culture; Cell isolation; Microscopy.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Animals, Newborn
Cell Culture Techniques / methods*
Cell Separation / methods
Cells, Cultured
Heart Ventricles / cytology*
Immunohistochemistry / methods*
Myocytes, Cardiac / cytology*
Rats