The establishment of the modified cultural technique of cardiomyocytes in human fetal hearts / 医学研究生学报

ABSTRACT

Objectives:This study was designed to establish the modified method of cardiomyocytes culture in human fetal hearts. Methods:The human fetal heart cells of ventricular muscle were isolated by 0.2% trypsin and 0.1% collagenase and cultured primarily and passaged in Iscove's modified Dulbecco's medium(IMDM) in vitro by means of differential attachment technique.The changes in morphology,ultrastructure,viability,immunocytochemistry antibody staining and immunofluorescence antibody staining of human fetal heart cells were studied in culture. Results:The ratio of viable cells was 99% identified by trypan blue staining.The ratio of attachment cells was 95% after 24 h in culture.The cultured human fetal heart cells were roundness shaped,rod shaped,shuttle shaped,ellipse shaped,star shaped and bifurcate shaped with spontaneous contractility.The myocardial actin and myoglobin are identified in the cultured cells by immunocytochemistry antibody staining and immunofluorescence antibody staining.The ultrastructure of cells was similar to that of the cardiac tissue in vivo by electron microscopy.Human fetal heart cells after 20 d of primary culturing and after 5 d of passaged culturing were growed well. Conclusions:The method for isolating and culturing human fetal heart cells is successful and reliable.This model provides an effective experimental mothod for studying the mechanism of myocardial injury.