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目的建立小鼠心房肌细胞分离技术,评价心房肌细胞的质量和电生理特性。方法通过改良Bustamante等人所建立的分离人心房肌细胞的方法,分离成年小鼠单个心房肌细胞。台盼蓝染色判定心房肌细胞活性。全细胞膜片钳技术记录细胞膜钾电流,证实细胞的电生理特性。结果分离出的小鼠心房肌细胞呈梭状、横纹清晰、两端边缘锐利、遮光性较强、细胞无明显搏动。全细胞膜片钳记录细胞膜钾电流(Ito,Ikur,Ik1),证实所分离的心房肌细胞具有典型的钾电流特征。结论 Bustamante法经改良后可以成功地获得适合做电生理需要的单个小鼠心房肌细胞,且易于操作,重现性好。
Objective To establish mouse atrial myocyte isolation technique to evaluate the quality and electrophysiological characteristics of atrial myocytes. Methods Single atrial myocytes of adult mice were isolated by modifying the method of isolating human atrial myocytes established by Bustamante et al. Trypan blue staining to determine the activity of atrial myocytes. Whole-cell patch clamp technique to record cell membrane potassium current, confirming the electrophysiological properties of cells. Results Isolated mouse atrial myocytes were spindle-shaped, clear stripes, sharp edges at both ends, strong shading, cells without significant beating. Whole-cell patch-clamp recording of membrane potassium currents (Ito, Ikur, Ik1) confirmed that the isolated atrial myocytes had typical K + current profiles. Conclusion The modified Bustamante method can successfully obtain a single mouse atrial myocyte that is suitable for electrophysiological needs and is easy to operate with good reproducibility.