单个人心房肌细胞急性分离技术及其细胞膜瞬时外向钾电流的记录  

Quick isolation of single atrial myocytes from the human heart and recording the transient outward potassium current by patch clamp

作　　者：邝素娟[1,2] 邓春玉[1,2] 杨慧[1,2] 陈怡静[1,2] 单志新[1,2] 林秋雄[1,2] 杨敏[1,2] 余细勇[1,2] 

机构地区：[1]广东省人民医院医学研究中心 [2]广东省医学科学院广东省心血管病研究所,广东广州510080

出　　处：《热带医学杂志》2013年第1期27-30,共4页Journal of Tropical Medicine

基　　金：国家自然科学基金(81070102);广东省自然科学基金(S2011020005911;10151008002000001);广东省科技计划项目(2010B080701090)

摘　　要：目的建立单个人心房肌细胞的急性分离技术,以满足检测人心肌细胞离子通道的需要。方法两步酶解法消化分离人心房肌细胞,采用膜片钳全细胞技术记录瞬时外向钾电流(Ito)。结果根据患者的性别、年龄及组织块的大小,选择适当的消化时间,分离得到结构清晰,横纹清楚,边缘整齐,少颗粒,舒展伸直,呈长杆状的单个心肌细胞;采用全细胞膜片钳技术能记录到瞬时外向钾电流。结论该方法能分离到形态和状态均良好的细胞,有利于进行单个人心房肌细胞膜片钳实验的研究。Objective To establish the method for quick isolation of single human atrial myocytes, which can provide a model for studying the ion channels in human cardiac ceils. Methods Single myocytes were isolated by two-step enzyme dissociation. Whole-cell patch clamp recording technique was used to record the transient outward potassium current （I_to）. Results According to the age, sex of the patients and the size of the tissue samples, we selected the appropriate time for digestion for isolation and obtained single ventricular ceils with clear structure, clear stripes, neat edge, less particles, stretch straight, and rod-shap. Transient outward potassium currents were recorded to use whole cell patch clamp technique. Conclusion We can get single atrial myocytes using this method, which can be used for the patch clamp experiments.

关 键 词：人心房肌细胞 分离 瞬时外向钾电流 膜片钳技术 

分 类 号：R331.38[医药卫生—人体生理学]