南方医科大学学报2013，Vol.33Issue(3)：397-400,4.DOI:10.3969/j.issn.1673-4254.2013.03.18
用于膜片钳研究的乳鼠窦房结细胞的分离与鉴定
A method for isolating and identifying newborn rat sino-atrial node cells for patch-clamp study
摘要
Abstract
Objective To compare the effects of enzyme digestion with differential attachment and enzyme digestion with differential attachment plus 5'-bromodeoxyuridine (5-BrdU) (conventional) method for isolating sino-atrial node cells from newborn rats for patch clamp studies. Methods The sino-atrial node tissue were removed from neonatal Wistar rats, digested and separated into single sino-atrial node cells with the above two methods. The obtained cells were observed for morphology, structure, beat frequency, degree of difficulty for patch clamp experiment and the success rate. The atrial cells isolated with enzyme digestion with differential attachment served as the control cells. Results The spindle-shaped cells constituted the greatest proportion of the cultured sino-atrial cells, and the cultured atrial cells showed mostly a triangular morphology. Both of the sino-atrial cells separated with the two methods contained mostly spindle-shaped cells, in which spontaneous action potential was recorded. The spindle-shaped cells isolated with enzyme digestion and differential attachment showed a greater viability with a higher success rate in patch clamp studies. Conclusion The sino-atrial node cells can be successfully isolated with enzyme digestion and differential attachment, which requires simple procedures and well preserves the cell viability for better performance in patch clamp experiment.关键词
窦房结细胞/分离/膜片钳/实验研究Key words
sino-atrial node cells/ isolation/ patch clamp/ experimental study引用本文复制引用
汪艳丽,刘如秀,刘宇,李泱..用于膜片钳研究的乳鼠窦房结细胞的分离与鉴定[J].南方医科大学学报,2013,33(3):397-400,4.基金项目
北京市自然科学基金(7102135) （7102135）
