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内耳I型纤维细胞培养物的特性鉴定与培养发展

Characterization and development of an inner ear type I fibrocyte cell culture.

作者信息

Gratton M A, Schulte B A, Hazen-Martin D J

机构信息

Department of Otolaryngology and Communicative Sciences, Medical University of South Carolina, Charleston 29425-4557, USA.

出版信息

Hear Res. 1996 Sep 15;99(1-2):71-8. doi: 10.1016/s0378-5955(96)00080-9.

Abstract

A method has been developed that allows successful maintenance of secondary cell cultures derived from explants of the cochlear lateral wall of young adult gerbils. The secondary cultures were characterized morphologically with light and transmission electron microscopy and immunocytochemically with protein markers specific to various lateral wall cell types. Structural studies revealed fusiform-shaped cells with a paucity of cytoplasm surrounding the nucleus and slender processes. The cells showed little evidence of intercellular contact even when confluent. The cultures were immunopositive for vimentin, carbonic anhydrase isozyme II, creatine kinase isozyme BB and smooth endoplasmic reticulum Ca-ATPase, but lacked reactivity for cytokeratins and Na,K-ATPase. The results indicate that the cultures are comprised of type I fibrocytes from the spiral ligament. These findings are the first to demonstrate that inner ear spiral ligament cells can be isolated and maintained in secondary culture while retaining many of their in vivo characteristics. Based upon their location and content of ion transport enzymes, type I fibrocytes are thought to be involved in the recycling of potassium from perilymph into the stria vascularis. The establishment of this cell line provides a means to analyze the role of spiral ligament fibrocytes in maintenance of inner ear homeostasis.

摘要

已开发出一种方法,可成功维持源自成年幼龄沙鼠耳蜗外侧壁外植体的二次细胞培养物。通过光学显微镜和透射电子显微镜对二次培养物进行形态学表征,并使用针对各种外侧壁细胞类型的特异性蛋白质标记进行免疫细胞化学分析。结构研究显示,细胞呈梭形,细胞核周围细胞质稀少,有细长的突起。即使细胞汇合,细胞间接触的迹象也很少。这些培养物对波形蛋白、碳酸酐酶同工酶II、肌酸激酶同工酶BB和平滑内质网钙ATP酶呈免疫阳性,但对细胞角蛋白和钠钾ATP酶无反应性。结果表明,这些培养物由来自螺旋韧带的I型纤维细胞组成。这些发现首次证明内耳螺旋韧带细胞可以在二次培养中分离并维持,同时保留其许多体内特征。基于其位置和离子转运酶的含量,I型纤维细胞被认为参与了钾从外淋巴回收到血管纹的过程。该细胞系的建立为分析螺旋韧带纤维细胞在内耳稳态维持中的作用提供了一种手段。

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