人眼Tenon's囊成纤维细胞原代培养的研究

In vitro culturing of human Tenon's capsule fibroblast cells

目的探讨体外培养原代人眼Tenon’s囊成纤维细胞的方法及其生长特性,为抗纤维化研究提供靶细胞模型。方法取翼状胬肉及斜视手术患者的Tenon’s囊组织,用组织块法培养原代成纤维细胞。用免疫荧光方法鉴定细胞。对细胞进行传代、冻存和复苏的观察。对复苏后的细胞行MTT法检测其活力并绘制生长曲线。结果人眼Tenon’s囊成纤维细胞可以在体外用组织块方法培养出来,呈典型的长梭形。免疫荧光鉴定细胞波形蛋白染色阳性。细胞多次传代后依然生长迅速,3 d即可长满瓶底。复苏后细胞2~6 d处生长对数期,增殖能力良好。结论人眼Tenon’s囊成纤维细胞体外生长状态良好,可以液氮冻存,复苏后细胞活力较强,可以用于抗纤维化增生的基础研究。

Objective To investigate in vitro culture method and growth characteristics of human Tenon’s capsule fibroblasts,a potential target cell model for the study on ocular anti-fibrosis treatment. Methods Human Tenon’s capsule tissue was collected from pterygium and strabismus surgery patients. Primary fibroblasts were harvested after tissue culturing and identified with immunocytochemical methods. Cells line passage,cryopreservation and recovery were observed. Cell activity and proliferation after recovery were examined with MTT. Results Human Tenon’s capsule fibroblasts could be cultured in vitro. Cells showed typical long spindle shapes,positively stained with vimentin. Cells grew rapidly after several passages and were able to cover the medium within 3 days. Logarithmic growth period usually occurred in 48 hours to 6 days after recovery. Conclusion Human Tenon ’s capsule fibroblasts could be cryopreserved in fluid nitrogen. After recovery these cells showed strong vitality and good in vitro growth. Cultured human Tenon’s capsule fibroblasts therefore could be used in the studies on ocular anti-fibrosis treatments.