豚鼠耳蜗外毛细胞的快速分离及活力观察

Quick isolation of cochlea outer hair cells in guinea pigs and their vitality

目的探讨豚鼠耳蜗外毛细胞(Outer Hair Cells,OHCs)的快速分离方法及其活力观察。方法采用机械与胰蛋白酶消化相结合的方法快速分离耳蜗OHCs,并在倒置显微镜下观察其活力。结果可快速分离出一定数量活性良好的耳蜗OHCs,其贴壁所需的时间较短,分离后静置7分钟即观察到贴壁。不同形态的OHCs在正常细胞外液中的存活时间不同,胞体较短的能存活3小时左右,胞体较长的能存活6～7小时。同时可观察到基底膜上其它几种细胞的形态。结论使用此方法可快速分离出单离的活性良好的耳蜗OHCs,这有助于我们深入研究其正常生理功能及某些病理状态下的功能及形态变化。

Objective To report a method of quick isolation of cochlea outer hair cells（OHC） in guinea pig and to examine their vitality.Methods OHCs were quickly isolated from the cochlea using a method of combined mechanical dissection and trypsine digestion.The cells＇ vitality was examined under an inverse microscope.Results A fair amount of living OHCs were obtained,which adhered in about 7 minutes after separation.OHCs of different morphologies showed different survival durations.Short cells survived for about 3 hours,while longer ones 6 to 7 hours.This method also allowed examination of a few other types of cells on the basilar membrane.Conclusion Our method provides a way to quickly separate single OHCs with good vitality,which are useful for in-depth study of OHC functions and their functional and morphological changes under certain pathological conditions.