Isolation of Ca2+-tolerant myocytes from adult rat heart.
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.
D000255:Adenosine Triphosphate / Q000032:analysis; D000818:Animals; D002118:Calcium / Q000494:pharmacology*; D002245:Carbon Dioxide / Q000378:metabolism; D002469:Cell Separation / Q000379:methods*; D003470:Culture Media; D007328:Insulin / Q000494:pharmacology; D008297:Male; D009206:Myocardium / Q000166:cytology* / Q000378:metabolism; D011188:Potassium / Q000494:pharmacology; D051381:Rats; D011919:Rats, Inbred Strains; D012710:Serum Albumin, Bovine / Q000494:pharmacology
D003470:Culture Media; D007328:Insulin; D002245:Carbon Dioxide; D012710:Serum Albumin, Bovine; D000255:Adenosine Triphosphate; D011188:Potassium; D002118:Calcium