原代乳鼠心肌细胞的培养方法

An method for primary culture of neonate rat cardiac myocytes

目的：探讨原代心肌细胞培养的条件和方法,培养纯度高、存活时间长的新生SD大鼠的心肌细胞。方法：用胰蛋白酶、胶原酶Ⅰ型和胶原酶Ⅱ型混合消化分离心肌组织,差速贴壁法和化学抑制法相结合纯化心肌细胞。结果：肌细胞24 h后贴壁生长,以梭形为主。48心肌细胞伸出伪足,立体感明显,折光性强。3-4 d后,相互接触交织成网,形成细胞簇,60%-80%的细胞开始搏动,搏动频率平均50次/min。四天后细胞搏动率可达到90%以上。培养至第40天,细胞停止搏动。结论：此种方法培养的心肌细胞纯度较高,存活时间长,是一种可靠的的原代乳鼠心肌细胞培养方法.

Objective∶To explore the conditions and approach to culture purified neonatal rat cardiocytes.Methods∶The typsinase,collagenaseⅠand collagenase Ⅱwas used to digest and isolate myocardial tissue,and then differential adhesion and chemical inhibition was was the way for purification.Results∶24 hours later,cells began to adhere and Spindle-shaped cell is main.48 hours later,cells stretch out the pseudopodia with Strong refractiion;3 ～ 4 days later,Pseudopodia are in contact to a network,cell clusters formed.60 %～80 % beating cardiomyocytes were observed and the beating frequency of cells was 50 beat s/ min after 3 days.Conclusion∶This method is an effective way to cultivate purified neonatal rat cardiocytes with long survival time.