犬心房肌细胞分离及其L型钙通道电流检测的研究

目的探讨一种稳定可靠的犬心房肌细胞的分离方法,提高膜片钳的实验成功率并检测L型钙通道电流。方法采用改良Langderoff灌流系统行主动脉逆行灌注,采用选择性冠状动脉插管的方法,获得单个心房肌细胞。在显微镜下观察细胞形态,并应用膜片钳全细胞模式记录L型钙通道电流。结果分离细胞存活率为70%～80%,复钙后细胞存活率为30%～40%,耐钙细胞形态呈杆状,折光性强,横纹清晰,并能稳定记录L型钙通道电流。结论改良Langderoff技术有助于稳定分离出存活率高、具有正常电生理特性的犬心房肌细胞,能用于心房肌细胞离子通道的研究。

A study on separation of canine atrial myocytes and detection of their L-type calcium channel current

Objective To explore a stable and reliable method of canine atrial myocytes separation for improving the success rate of patch-clamp techniques and detecting the L-type calcium channel current.Methods Modified Langderoff perfusion system was adopted to perform retrograde aortic perfusion and selective coronary artery catheterization was used to obtain single atrial myotcytes.Microscope was employed to observe the cellular morphology and the L-type calcium channel current was measured by whole-cell patch-clamp recording.Results The survival rate of separated cells was 70% to 80%,and it was 30% to 40% after recalcification.A stable L-type calcium channel current could be recorded in calcium-tolerant cells which was rod-shaped with good refraction and clear horizontal stripe.Conclusion Modified Langderoff technique contribute to stable separation of canine atrial myocytes with high survival rate and normal electrophysiological properties,and can be used in the research of the iron channel of atrial myocytes.