乳鼠心肌细胞原代培养方法  

Methods of primary culturing myocardial cells of neonatal rat

作　　者：刘嘉祺[1] 孙云云 

机构地区：[1]牡丹江医学院,黑龙江牡丹江157011 [2]威海市立第二医院,山东威海264200

出　　处：《微量元素与健康研究》2012年第4期4-6,共3页Studies of Trace Elements and Health

摘　　要：目的:探讨新生乳鼠心肌细胞原代培养的条件和方法,培养纯度高、存活时间长的心肌细胞。方法:参照Simpson等的方法加以改进,采用含0.05%EDTA的胰蛋白酶多次消化心肌,差速贴壁1.5 h纯化心肌细胞进行培养,培养过程在37℃、5%CO2条件下进行。用台盼蓝染色法检测培养细胞的存活率,倒置显微镜下观察细胞形态变化,记录3 d、6 d、9 d、12d心肌细胞搏动频率,测定细胞活力。结果:原代培养乳鼠心肌细胞生长良好,培养3 d、6 d、9 d、12 d时,用台盼蓝染色法检测心肌细胞的存活率均大于96%,培养12 d时仍为97.02%。在倒置显微镜下观察,培养48 h左右的细胞之间相互连接的并有细胞搏动。培养6 d、9 d、12 d的细胞与培养3 d的相比,活力增强,差异显著(P<0.05),从6 d开始,细胞活力并未显著增强,两两比较均无统计学差异(P>0.05)。结论:应用此方法培养的心肌细胞纯度高,存活时间长,是一种稳定、可靠的原代心肌细胞培养方法。Objective：To explore the conditions and methods of primary culturing myoeardia! cells of neonatal rat, and to culture high purity and long time surviving myocardial ceils. Methods： In accordance with Simpson, myocardial tissue was digested with trypsin of 0.05 % EDTA repeatedly, myocardial cells were purified with differential adherence method for one and half an hours at 37 ℃ in humid air with 5 % CO2. Cell survival rate was detected with Trypan blue staining and cell morphologic change was observed under an inverted microscope. To record beating rate of cardiac myocyte after 3 days, 6 days, 9 days and 12 days. Results： Primary cultured myocardial cells of neonatal rat grew well, survival rate of myocardial cells was more than 96 % after cells were cultured for 3 days, 6 days, 9 days and 12days. And it was also 97.02% after 12days. Ceils were observed under an inverted microscope,we found that ceils conjoined to each other and beat after 24 hours. Cell vigor after 6 days, 9 days and 12 days was stronger than that after 3 days, and discrepancy was significant（P 〈 0.05 ）. From the sixth day, cell vigor was no reinforcement notablely, and it was no statistical discrepancy to each other （P 〉 0.05 ）. Conclusion： Myocarctial cells cultured with this method were high purity and survival for long time, so it waa a stable and reliable method to culture primary myocardial cells.

关 键 词：乳鼠 原代培养 心肌细胞 

分 类 号：Q331.38[生物学—遗传学]