用于钙信号研究的小鼠心室肌细胞急性分离的优化
Medical Research and Education（2016）
Abstract
目的：建立一种可快速获取生理状态良好的用于钙信号研究的小鼠心室肌细胞的方法，同时比较两种细胞保存液对心室肌细胞钙信号的影响。方法取成年小鼠心脏，用改进的 Langendorff 心脏灌流系统行主动脉逆行灌流，利用酶联合法消化，取得心室肌细胞置于两种不同的细胞保存液中，在显微镜下观察并检测其状态。采用共聚焦显微镜观测细胞内钙信号，记录钙火花。结果该方法可以在较短的时间内得到65％以上的长杆状心室肌细胞，细胞能够正常存活12～24 h，并且可以立即用于钙火花观测实验研究。比较两种保存液，细胞保存液 A中富含高钾离子，细胞保存液B中含有正常的胞外钙离子浓度，观察到的钙火花细胞保存液B中比A液的频率高、幅度大，在达峰时间上比A液的时间短。结论通过严格控制各项实验条件，能够成功得到存活率高、纯度高、耐钙的可用于钙火花观测的小鼠心室肌细胞。细胞保存液中的钾离子含量少，并有适量的钙离子有助于观测钙火花的发生。
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