不同用途的心室肌细胞的分离

Improvement of cardiomyocyte isolation for different purposes

目的 :建立适合于不同用途的心室肌细胞的酶解分离方法。方法 :恒流 Langendorff灌流 ,I型胶原酶酶解分离 ,应用膜片箝技术记录离子电流、双波长显微荧光光度术检测细胞内游离钙离子浓度及视频跟踪系统测定心肌单细胞收缩。结果 :采用不同分离技术参数获得的心肌细胞能满足上述三种实验条件下的要求。酶解分离的单个心室肌细胞静息电位为 (- 74 .0 6± 4 .5 4 ) m V,记录到心室肌细胞典型的钾、钠、钙离子电流和稳定的心室肌细胞内游离钙比值 ,心室肌单细胞收缩特性稳定。结论 :通过调整分离技术参数获得的心室肌细胞性能稳定 ,具有正常的电 -机械生理特性和耐钙性。

Objective: To improve the methods of isolating single ventricular myocytes for purposes of different experiments. Methods: The cardiac ventricular myocytes were isolated enzymatically by Langendorff perfusion technique at constant flow rate. The patch clamp whole-cell recording, a spectrofluorometric method and a video-tracking system were used to verify the basic electrophysiological properties, intracellular calcium transient and cell contraction of the single myocytes. Results: The resting membrance potential of the myocyte was (-74.06± 4.54)mV. The whole-cell sodium, potassium and calcium currents, intracellular calcium transients and contractile properties were stable and typical. Conclusion: The myocytes obtained by adjusting perfusion parameters are well suited for electrophysiological recording, intracellular calcium and single cell contraction measurements.