The Establishment of the Apoptotic Model of Cultured Human Adult Cardiomyocytes / 中国医师杂志

ABSTRACT

Objective To establish a model for hypoxia-induced apoptosis of human adult cardiomyocytes in vitro. Methods The human adult heart cells of ventricular muscle were isolated by the digestion of trypsin and collagenase, cultured in Iscove's modified Dulbecco's medium (IMDM), and purified by means of differential attachment technique, adding Brdu and slicing fibroblasts by sterile needle. After 4 to 6 days, the cardiomyocytes were cultured in an incubator containing 3% O 2, 92% N 2 and 5% CO 2 for 6, 12, 24 and 48h to form hypoxia injury for cells. Morphological changes of apoptotic cardiomyocytes were measured by invertion microscopy and electronic microscopy. Apoptotic rate was measured by TUNEL staining. Results The cardiomyocytes occurred opoptosis after hypoxia injury, apoptotic rate of which at 6, 12, 24 and 48h after hypoxia was (3 3?0 9)%, (8 3?1 8)%, (16 1?2 6)% and (19 4?2 3)% respectively. Conclusion The apoptotic rate of cardiomyocytes increased with time of hypoxia lengthened. The model for hypoxia-induced apoptosis of human adult cardiomyocytes was successful and reliable.