摘要
目的:介绍1种改进的成年SD大鼠心房肌细胞分离方法。方法:SD大鼠麻醉后,快速取出心脏,将主动脉结扎在Langendorff灌流装置上,无钙台式液灌流5 min后换用酶液灌流10～12 min,剪取心耳组织于KB液中剪碎并吹打,200目细胞筛网过滤,将细胞滤液500 r离心2 min,梯度复钙后将细胞悬液置于4℃冰箱中静置1 h,应用全细胞膜片钳技术记录大鼠心房肌细胞瞬时外向钾电流(Ito)、快钠电流(INa)以及L型钙电流(ICa-L)。结果:本方法分离出的心房肌细胞存活率高,结构完整,具有良好的耐钙性,可成功记录出Ito、INa、ICa-L。结论:改进的实验方法可获得大量耐钙心肌细胞,并具有正常的电生理特性。
Objective：To introduce a modified isolation method for acquiring rat atrial myocytes.Methods：The heart of adult sprague-dawley（SD） rat was rapidly excised after successful anesthesia and mounted on a Langendorff apparatus.It was perfused retrogradely via the aorta for five minutes with Ca2＋-free Tyrode＇s solution and then perfused with collagenase dissolved in Tyrode＇s solution for ten to twelve minutes,and then the atrial appendage was sheared and stored in a kraftbruhe（KB） solution.Thereafter tissue was minced and filtrated.After being centrifuged at 500 r for 2 min,the isolated myocytes were performed gradient calcium reintroduction and allowed to pellet via gravity for 1 hour in refrigerator,then the transient outward potassium current,sodium current and L-tpye calium current were recorded by the use of patch clamp whole cell recording technique.Results：The survival rate of atrial myocytes was high and the morphous and construction of atrial myocytes were fine and the property of calcium-tolerance was good.The transient outward potassium current,sodium current and L-tpye calium current were recorded successfully.Conclusion：The modified isolation method can produce large numbers of single calcium-tolerance myocardial cells and normal electrophysiological properties.
出处
《重庆医科大学学报》
CAS
CSCD
北大核心
2011年第8期910-912,共3页
Journal of Chongqing Medical University
基金
国家自然科学基金资助项目(编号:30871075)
关键词
膜片钳
心肌细胞
瞬时外向钾电流
钠电流
L型钙电流
细胞分离
patch clamp
atrial myocytes
transient outward potassium current
sodium current
L-type calium current
cell isolation
