豚鼠耳蜗单离Deiters细胞分离方法及活性判定标准

Method of Isolating Deiters Cell from Guinea Pig Cochlea and Criterion of Cell Judgement

目的 探讨分离单离Deiters细胞的方法及活性判定标准。方法 分别用胶原酶、木瓜蛋白酶和胰蛋白酶消化基底膜后结合机械吹打分离出Deiters细胞 ,观察并计数。结果 单离的Deiters细胞呈豆点状 ,分胞体、柄、指突三部分 ,平均每双侧耳可分离到 2 0～ 32个单离的Deiters细胞。活性标准 :细胞膜光滑 ,双折射现象存在 ,细胞核位置正常 ,指突完整 ,无呈布朗运动的颗粒。结论 三种常用的酶孵育后结合吹打法均可以分离出相当数量且具有活性保存较好的单离Deiters细胞 ;

Objective To probe the method of dissociating isolated Deiters cells of cochlear and its judging criterion for living Deiters cells.Methods Deiters cells were dissociated from guinea pig cochlea separately by collagenase,papain,trysin digestion followed by trituation.After that ,the Deiters cells were observed and counted.Results Isolated Deiters cell looked like a comma, which was divided into cell body, stem and Deiters phalanx. About 20 to 32 isolated Deiters cells were obtained from every two cochleae.The membrane was smooth and the birefringence existed; the position of the nucleus was normal; full phalanx and no particles with Brownian movement.Conclusion Substantial sum of Deiters cell with sound vitality can be obtained by all three enzymes digestion followed trituation;The criterion used is reliable.