Isolation of Ca2+-tolerant myocytes from adult rat heart

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Abstract
A procedure for the isolation of myocytes from adult rat hearts is described. It is based on successive treatments with Ca2+-free medium, disaggregating enzymes (collagenase and hyaluronidase) and mechanical agitation. Several recent isolation methods were compared and their best features were combined, together with some original modifications. A good yield of high purity myocytes with excellent morphological and functional integrity was obtained. The cells are tolerant to physiological concentrations of Ca2+. Cellular levels of ATP, Na+ and K+ are close to those in intact hearts, and glucose oxidation rates and succinate exclusion are also close to normal. These characteristics are maintained for periods over 1 h.