Key result
A protocol using collagenase and protease perfusion under low calcium conditions yields adult rat heart myocytes that remain viable in culture for up to four days.
This protocol provides a reliable method for isolating and culturing viable adult rat heart myocytes for up to four days for use in cardiovascular research.
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Supports short-term adult rat myocyte culture; extends preclinical tools but leaves translation to humans open.
Cultured primary adult rodent heart cells are an important model system for cardiovascular research. Nevertheless, establishment of robust, viable cultured adult myocytes can be a technically challenging, rate-limiting step for many researchers. Here we described a protocol to obtain a high yield of adult rat heart myocytes that remain viable in culture for several days. The heart is isolated and perfused with collagenase and protease under low Ca2+ conditions to recover single myocytes. Ca2+-tolerant cells are obtained by stepwise increases in extracellular Ca2+ concentration in three subsequent wash steps. Cells are filtered, resuspended in culture medium, and plated on laminin coated slips. Cultured myocytes obtained using this protocol are viable for up to four days and are suitable for most experiments including electrophysiology, biochemistry, imaging and molecular biology.
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Xu et al. (2009) studied Cardiovascular research model. Protocol for primary culture of adult rat heart myocytes was evaluated on Viability of cultured adult rat heart myocytes. A protocol using collagenase and protease perfusion under low calcium conditions yields adult rat heart myocytes that remain viable in culture for up to four days.
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