成年大鼠心肌细胞培养方法的建立和形态学观察

Morphology Study and Isolated and Cultured Method of Adult Myocytes

为探索稳定的成年大鼠心肌细胞的分离和培养方法 ,应用生物酶 ( 5g/L胰蛋白酶及 1 g/L胶原酶 )灌注法分离成年大鼠心肌细胞 ,用形态学及台盼蓝染色法鉴定分离的心肌细胞 ,并在光镜和电镜下观察和摄像记录。结果 :心肌细胞的存活率为 91 .7% ;存活的心肌细胞静止地贴在培养板底上 ,细胞完整 ,呈杆状 ,长宽比约为 4～ 6∶1。结果提示 :该方法是比较理想的心肌细胞培养法。

In order to build method of a stable isolation and culture of adult myocytes, adult rat myocytes were isolated with bioenzymes (including 5 g/L trypsin and 1 g/L colleagase) perfusing tissue through acceding aorta of the heart. Calcium tolerant myocytes were harvested, purifyed and cultured with DMEM culture medium . The myocytes quality was evaluated with morphology and trypan blue dye. States of myocytes growing in culture were observed with optical microscope and electron microscope photography. Result: Survival rate of aduld rat myocytes was 91.7%, survival myocytes were motionlessly attached to the bottom of culture plates, they were intact and rod shaped, their length/width ratio was 4～6: 1. Conclusion: The method of isolation and culture of adult rat myocytes was successful.