适于膜片钳实验的大鼠心肌细胞分离  

Isolation of cardiomyocytesfrom rats for patch clamp studies

作　　者：许正新[1] 郑雪[1] 祝丽[1] 朱磊[1] 

机构地区：[1]扬州大学医学院,江苏扬州225001

出　　处：《临床医药文献电子杂志》2015年第33期6761-6762,6765,共3页Electronic Journal of Clinical Medical Literature

基　　金：国家自然科学基金资助(NO.31071171)

摘　　要：目的 对酶解分离大鼠心肌细胞的条件进行探索并优化。方法 采用Langendorff装置,酶解恒温灌流消化及梯度复钙等方法对大鼠心肌细胞进行分离和处理。结果 当钙离子浓度为30μm、胶原酶含量0.4 g/L,分离时间为20~30 min的条件下,可分离得到大量边缘光滑、完整,横纹清晰,表面无空泡,长杆状的心肌细胞,其比例约为60%~80%,将细胞室温静置2~6 h,采取梯度复钙法对所得细胞进行复钙处理,虽有部分细胞出现收缩、死亡,但仍然可得到形态好、耐钙、静止无收缩,容易形成高阻封接、易破膜的心肌细胞,其比例约为40%~50%。结论：本实验探索出的心肌细胞分离方法过程简便,结果稳定,可用于后续膜片钳实验。Objective Explore and optimize the myocardial cell separation by enzymolysis conditions. Methods The cardic myocytes were isolated enzymatically by langendorrf perfusion and recalcification step by step. Results With the calcium ion concentration of 30 μM, collagen enzyme content of 0.4 g/L, the separation time of 20-30 min conditions, a large amount of rat cells with smooth and complete edge were acquired, they were in long rod shaps and no cavity surface and their cross grain were clear.The initial surval rate was 60%~80% and surval rate after recalcification step by step was 40%~50%, but the surval myocytes were still in good shaps, calcium tolerance,still and no shrinkage, easy to form a high resistance sealing though the part of cell death. Conclusion The introduced isolation method is easy with good stability which are applicable for the study on the experiment of patch calmp.

关 键 词：大鼠 心肌细胞 细胞分离 膜片钳 

分 类 号：R332[医药卫生—人体生理学]