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应用胰蛋白酶分离豚鼠耳蜗外毛细胞
Isolation of Cochlear Outer Hair Cells from Guinea Pigs by Trypsin
【摘要】 目的 :采用胰蛋白酶孵育后机械分离法以获得单离的豚鼠耳蜗外毛细胞。方法 :豚鼠被迅速断头处死 ,暴露耳蜗 ,在解剖显微镜下去除耳蜗侧壁 ,切取第三、四回耳蜗 ,经胰蛋白酶消化并轻轻吹打 ,以获得单离的豚鼠耳蜗外毛细胞。静置后在倒置相差显微镜下观察。结果 :本法每侧耳分离出活性好的外毛细胞约 4 0~ 90个 ,能存活 5小时左右。结论 :此技术能提供足量的单离毛细胞 ,用于细胞膜的电生理性质等研究
【Abstract】 Objective:To obtain isolated outer hair cells from guinea pigs,by trypsin digestion and mechanical agitation.Methods:Guinea pig was decapitated and the temporal bone was quickly removed.Then the lateral wall of the cochlea was removed.The 3 rd and the 4 th turn of the organ of Corti were dissected out under a dissection microscope.Single cells were obtained by digestion with trypsin and gentle mechanical agitation.Results:The isolated outer hair cells were observed under inverse microscope.Forty to fifty single outer hair cells were obtain from each ear and most of them could survive for more than 5 hours.Conclusion:This technique can provide sufficient numbers of solitary OHCs for electrophysiological studies of the cell’s membrane properties.
【Key words】 outer hair cells; isolate; trypsinThe Study on Physical Toxicity of Methamphetamine;
- 【文献出处】 华西医学 ,West China Medical Journal , 编辑部邮箱 ,2003年01期
- 【分类号】R764
- 【被引频次】14
- 【下载频次】122