应用胰蛋白酶分离豚鼠耳蜗内毛细胞

Isolation of Cochlear Inner Hair Cells from Guinea Pigs by Trypsin

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内毛细胞。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 ,经胰蛋白酶消化并轻轻吹打 ,以获得单离的豚鼠耳蜗内毛细胞。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内毛细胞约 5～ 2 0个 ,多数分离的内毛细胞能存活 5 h左右。结论 此技术能提供足量的单离内毛细胞 。

Objective To obtain isolated inner hair cells from guinea pigs by trypsin digestion and mechanical trituration. Methods The guinea pig was decapitated and the temporal bone was quickly removed. Then the lateral wall of the cochlea was removed. The organ of Corti was dissected out under a dissection microscope. Single hair cells were obtained by digestion with trypsin and gentle mechanical trituration. Results The isolated inner hair cells were observed under inverse microscope. Five to twenty single outer hair cells were obtained from each ear and most of them could survive for about 5 hours. Conclusion This technique can provide sufficient numbers of solitary inner hair cells for studies on the electrophysiological properties of the cells.