适用于膜片钳研究的新生大鼠骨骼肌细胞培养

THE CULTURE OF SKELETAL MUSCLECELLS SUITABLEFOR PATCH-CLAMPING STUDY FROM NEONATALRATS

采用低浓度胰蛋白酶，较低温度下长时间搅拌及粗口径滴管分次吹打等改进方法进行新生大鼠骨骼肌细胞分散培养，能够得到较高的细胞产率。经１０ｎｍｏｌ／Ｌ秋水仙碱处理３ｄ后可得到肌球，能够适合于进行膜片钳记录，并不改变肌细胞的电生理学特性，肌球膜上有正常的Ｎａ￣＋、Ｋ￣＋和Ｎ型乙酰胆碱受体离子通道活动。本方法提供了一个较好的适用于肌细胞电生理学研究的模型。

Low concentration of trypsin,prolonged time of dissociation with stirring at low tempera-ture and gentle trituration with large-diameter pipettes were adopted to culture skeletal musclecells from neonatal rats.The rate of live cells was high.Three days after treatment with 10nmol/L colchicine, spherical myoballs formed spontaneously from myotubes.Myoballs couldbe used in patch-clamping experiments especially in whole-cell-recording mode,The colchicinehad no apparent effect on the electrophysiological properties of the muscle cells.There were nor-mal Na￣ +,K￣and N type ACh receptor ion channels in the membrane of myoball.The cultureof myoballs could be a suitable preparation for patch,clamping studies on cultured rat skeletalmuscles.