成年大鼠心肌细胞急性分离方法的探讨  

A method of acute isolation of adult rat ventricular cardiomyocytes

作　　者：张琳[1] 纵静[2] 张庆涛[3] 

机构地区：[1]徐州医学院临床学院,江苏徐州221002 [2]徐州医学院附属医院,江苏徐州221002 [3]徐州医学院附属第三医院,江苏徐州221003

出　　处：《徐州医学院学报》2015年第10期696-699,共4页Acta Academiae Medicinae Xuzhou

基　　金：国家自然科学基金（81400178）;徐州市科技计划项目（XM138062）

摘　　要：目的：探讨成年大鼠心肌细胞急性分离过程中各种影响因素，建立稳定、高效、易重复的成年大鼠心肌细胞急性分离方法。方法成年SD大鼠麻醉后迅速取出心脏，悬挂在Langendorff装置上，用Ⅱ型胶原酶经升主动脉逆行灌流法分离大鼠心肌细胞，其间控制各种影响因素。结果分离即刻活细胞产量（3．2±0．4）×107，长杆状、横纹清晰；台盼蓝染色阴性的杆状细胞比率（86．7±5．3）％，经培养细胞可存活7天以上。结论通过调节和控制心肌细胞急性分离过程中的各种影响因素，可以获得高产量、高活性的心肌细胞。Objective To explore the factors involved in the separation of adult rat ventricular cardiomyocytes in or-der to establish a stable, efficient and repeatable method.Methods Adult rat hearts were quickly removed under anaes-thesia and hung on the Langendofff apparatus.Then, cardiomyocytes were isolated from SD rats by collagenase type Ⅱperfusion.The morphology of cardiomyocytes was observed by optical microscopy.The viability of cardiomyocytes was e-valuated by trypan blue exclusion test.Results After isolation, the number of viable cells per rat was （3.2 ±0.4）× 107, with a negative rate of （86.7 ±5.3）% after trypan blue staining.The cardiomyocytes could survive more than 7 days.Conclusion Adult rat cardiomyocytes can be isolated more efficiently through regulation of all factors involved in the isolation.

关 键 词：成年大鼠 心肌细胞 急性分离 影响因素 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]