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[从豚鼠耳蜗中分离毛细胞的技术]

[Techniques for isolating hair cells from guinea pig cochlea].

作者信息

Su Z

机构信息

PLA General Hospital, Beijing.

出版信息

Zhonghua Er Bi Yan Hou Ke Za Zhi. 1992;27(3):133-5, 189.

PMID:1419187
Abstract

Isolated live hair cells are important models for studying the electrophysiology, pathology and pharmacology of the hair cell. Using mechanical isolation method after papain treatment 70 +/- 27 hair cells, including 0 to 4 inner hair cells, could be obtained from each cochlea of 4 guinea pigs. The criteria for a good viability of isolated cochlear hair cells were: 1. a smooth hair cell membrane; 2. hair cells not swollen; 3. the nucleus in the normal position; 4. the cytoplasm in a state of semitransparency with a halo at the periphery (birefringence) and 5. no Brownian movement of the organelles within the cytoplasm. With short-term culturing at room temperature, approximately 90% of the isolated hair cells retained a good viability at the end of two hours. Subsequently the hair cells gradually degenerated but still, at the end of five hours, about 40% of them appeared intact. The degeneration patterns have been carefully observed and described.

摘要

分离出的活毛细胞是研究毛细胞电生理学、病理学和药理学的重要模型。采用木瓜蛋白酶处理后的机械分离方法,从4只豚鼠的每个耳蜗中可获得70±27个毛细胞,其中包括0至4个内毛细胞。分离出的耳蜗毛细胞具有良好活力的标准为:1. 毛细胞膜光滑;2. 毛细胞未肿胀;3. 细胞核位于正常位置;4. 细胞质呈半透明状态,周边有光晕(双折射);5. 细胞质内细胞器无布朗运动。在室温下进行短期培养,约90%的分离毛细胞在两小时结束时仍保持良好活力。随后毛细胞逐渐退化,但在五小时结束时,仍有约40%的毛细胞看起来完好无损。已对退化模式进行了仔细观察和描述。

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