豚鼠血管纹缘细胞的原代培养

Primary culture of strial marginal cells of guinea pig cochlea

目的:建立豚鼠内耳血管纹缘细胞的体外培养系统,为研究缘细胞的功能提供良好的材料。方法:采用移植培养技术,将活体分离的血管纹组织块接种于无菌塑料培养皿中,于5％CO2 37℃恒温箱内培养,每周换液2次,观察细胞生长情况。利用免疫组织化学技术,将抗细胞角蛋白抗体和抗波形蛋白抗体用于检测培养细胞。制作透射电镜标本观察培养细胞的超微结构。结果:培养细胞成功生长4周,具有多角形细胞和梭形细胞两种不同形态,免疫组织化学反应显示角蛋白、波形蛋白染色于多角形细胞胞浆中,梭形细胞仅波形蛋白呈阳性反应。透射电镜示多角形细胞具有紧密接合和桥粒等上皮细胞的特征。结论:采用移植块培养技术,成功建立豚鼠内耳血管纹缘细胞的原代培养方法,为研究缘细胞的功能提供了合适的细胞模型。

Objective.'For further study of the function of marginal cells, primary cultures of marginal cells of guinea pig were established. Method:Tissue of stria vascularis was transferred from the cochlea to a cell-cultured dish using the explants culture technique. Explants were cultivated at 37C in a 5% CO2 incubator and examined daily by interted microscopy. The culture medium was changed twice a week. Antibodies of cytokeratin and vim-entin were used to examine the cultured cells by immunohistochemical methods. The ultrastructure of the cultured cells was observed by making the electron microscopy specimens. Result:Explants were successfully cultivated for 4 weeks. Two different cells, hexagonal-shaped cells and spindle-shaped cells were observed. Cytokeratin and vim-entin staining occurred in cytoplasm of hexagonal-shaped cells, whereas spindle-shaped cells gave a negative cytokeratin staining but a positive vimentin staining. Electron microscopy of the explants revealed that hexagonal-shaped cells have the epithelial morphologic characteristics such as tight junctions and desmosomes. Conclusion.-Primary culture of marginal cells by explants culture technique provides an optimal model for investigation of the function of the marginal cells.