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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: To establish an in vitro culture system of guinea pig inner marginal cells, providing a good material for studying the function of marginal cells. Methods: Transplantation culture technique was used to inoculate the biopsied vascular tissue into sterile plastic petri dishes and cultured in a 5% CO2 incubator at 37 ℃ for 2 times a week to observe the cell growth. Anti-cytokeratin antibodies and anti-vimentin antibodies were used to detect cultured cells using immunohistochemistry. Transmission electron microscopy was used to observe the ultrastructure of cultured cells. RESULTS: The cultured cells grew successfully for 4 weeks, with two different morphologies of polygonal cells and spindle cells. Immunohistochemical staining showed that keratin and vimentin were stained in the cytoplasm of polygonal cells, and the spindle cells were only vimentin-positive . Transmission electron microscopy revealed that polygonal cells have features of tight junctions and epithelial cells such as desmosomes. CONCLUSION: The primary cultivation method of the vascular marginal cells in the inner ear of guinea pigs is successfully established by the transplantation block culture technique, which provides a suitable cell model for studying the function of marginal cells.