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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

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【作者】 唐玥玓张瑞林鲜均明

【Author】 TANG Yue-di,ZHANG Rui-lin,XIAN Jun-ming(Department of Otolaryngology-Head &Neck Surgery,West China Hospital,Sichuan University,Chengdu 610041,China)

【机构】 四川大学华西医院耳鼻咽喉科学研究室四川省成都市第二人民医院耳鼻咽喉科四川大学华西医院耳鼻咽喉科学研究室 四川成都610041四川成都610017四川成都610041

【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.

  • 【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
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