摘要
目的 :探讨豚鼠耳蜗部分外支持细胞 (Deiter细胞、Hensen细胞和外柱细胞 )的分离方法 ,并建立细胞活性的鉴别标准。方法 :选取健康杂色豚鼠 5只 ,解剖出耳蜗基底膜 ,采用酶解加机械吹打法分离Deiter细胞、Hensen细胞和外柱细胞。结果 :可以获得较多数量、活性良好、长短不一的Deiter细胞和Hensen细胞 ,8h内细胞活性较好 ;但外柱细胞数量较少 ,存活时间较短。评价Deiter细胞、Hensen细胞和外柱细胞的活性标准 :①细胞膜无膨胀或扭曲 ;②细胞核无肿胀、移位 ;③在相差显微镜下细胞呈半透明状态 ,存在双折射现象 ;④细胞内无呈布朗运动的颗粒。结论 :熟悉耳蜗的解剖特性、分离出完整的基底膜 ,增加酶的浓度和加大机械吹打力度是获取活性良好的Deiter细胞、Hensen细胞和外柱细胞的关键 ;评价外毛细胞活性的 4项标准同样可用于判断Deiter细胞。
Objective:To investigate the method of isolating partial outer supporting cells (Deiters' cells, Hensen cells and Outer pillar cells) from guinea-pig cochlea and to try to establish the morphological criteria for these outer supporting cells viability.Method:The basal membrane was dissected from the pigmental guinea pig cochlea, and th incubated with enzyme ( 0.5 mg/ml),then mechanically trituated by micropipetee.Result:A large number of living Deiters' cells and Hensen cells could be obtained from the guinea pig cochlear which kept a good condition in the next eight hours. However, the isolated outer pillar cells, with a more vulnerable viability, had a low yield and degenerated in the followed five hours.Conclusion:The key to success in isolating outer supporting cells, mainly Deiters' cells and Hensen's cells, from the guinea pig cochlea is to familiarize with the anatomical characteristics of the cochlea and to get the intact organ of Corti, as well as to increase the concentration of enzyme. Moreever, the later four criteria for outer hair cells viability are also fit well for these isolated Deiters' cells, Hensen's cells and Outer pillar cells.
出处
《临床耳鼻咽喉科杂志》
CAS
CSCD
北大核心
2004年第1期38-40,共3页
Journal of Clinical Otorhinolaryngology
