An improved method for isolating Ca2+-resistant myocytes from the adult rat heart.
Maintaining viability in cardiac myocytes isolated from adult rats using collagenase is difficult in Ca-containing media due to cell damage that occurs on reintroduction of Ca after perfusing the heart with the Ca-free medium needed to isolate myocytes with collagenase. Recently it has been proposed that Ca-free perfusion of isolated rabbit interventricular septa leads to cellular Na overload which, on reintroducing Ca, produces influx of toxic concentrations of Ca due to the Na/Ca exchange mechanism in the sarcolemma. We have found that replacing a portion of the 118 mM NaCl in the Ca-free perfusion medium with 69 mM LiCl dramatically increased the proportion of Ca-tolerant cardiac myocytes isolated from adult male rats with collagenase. Myocyte viability was maintained over a four hour period of incubation at 37 degrees C in 1 mM Ca.
Khetarpal
V K
VK
Department of Pharmacology, Thomas Jefferson University, Philadelphia, PA 19107.
Kocsis
J J
JJ
Weiss
A J
AJ
D000818:Animals; D002118:Calcium / Q000378:metabolism* / Q000494:pharmacology; D003584:Cytological Techniques; D009200:Myocardial Contraction / Q000187:drug effects; D009206:Myocardium / Q000166:cytology*; D051381:Rats; D014343:Trypan Blue
D014343:Trypan Blue; D002118:Calcium