介绍一种成年兔心肌细胞的分离与培养方法

Isolation and Culture of Adult Rabbit Cardiomyocytes

详细介绍了成年兔心肌细胞的分离方法 (胶原酶二步灌流法 )以及分离后的心肌细胞的培养 .根据实践经验 ,强调了灌流失败的主要原因是灌流管道末端插入过深以及胶原酶的用量不适 .同时提出在整个灌流和细胞制备过程中 ,要严格控制灌流液及胶原酶液的pH为 7 35～ 7 4 5 ,保持灌流液温度在 37℃ ,以减少细胞的损伤和死亡 .还提醒读者 。

An isolation method of adult rabbit cardiomyocytes (two steps collagenase perfusion method) and cardiomyocyte culture after isolation were introduced in detail. According to experience from practice, it is emphasized that main causes of fault is too deep insertion of the perfusion tube end and collagenase dosage is improper. It is noted that the pH of perfusion and collagenase fluids should be kept between 7 35 and 7 45, and the temperature of perfusion fluid should be kept at 37℃, so that the damaged and death cell numbers can be reduced. It is reminded that the same kind of colllagenase may have evidently different effects on the dosage and the isolation results because of different productive batch.