An improved method of isolation of rat cardiac myocytes / 中国药理学通报

ABSTRACT

Aim To introduce an improved method of Langendorff perfusion of the isolated rat heart that is easy to determine the termination of digestion.Methods Hearts were excised quickly from anesthetized SD rats.After the perfusate was free of blood,the solution was changed to perfusion buffer(0.6% Collagenase B,0.6% BSA,30 ?mol?L-1 Ca2+ in Tyrode solution)at 37℃.Hearts were isolated by traditional and improved Langendorff perfusion.Individual myocardial cell was measured by video-based motion edge-detection system(IonOptix,USA).Results One group of heart was digested by traditional Langendorff perfusion for 13~16 min.The termination of digestion could not be judged properly by this method.The nature and quality of the cardiocytes were various.The cardiocytes could not keep their survival and viability when exposed to Tyrode Solution with 1.8 mmol?L-1 Ca2+.Another group was digested by improved Langendorff perfusion.More than 80% survival cardiac ventricle myocytes could be obtained by improved Langendorff perfusion.Moreover,about 50% cardiac myocytes exposed to Tyrode solution with 1.8 mmol?L-1 Ca2+ could retain rod-shaped and be used to contraction research.The contraction of cardiocyte was stabile within 1 000 s.Conclusion Cardiac myocytes disassociated from improved Langendorff perfusion can be used in these studies of detecting contraction and relaxation.This method is economical and easy to control.The beginners are able to acquire the technological method over a short-term practice.