兔和牛角膜上皮、基质及内皮细胞体外培养和增殖的研究

Cytobiological study of three corneal cell types in vitro

目的:研究兔和牛角膜上皮、基质及内皮细胞等3种细胞体外培养的生物学特性,探索和改进这3种细胞体外培养的方法,为组织工程角膜奠定基础。方法:采用消化法分离培养兔和牛角膜上皮、内皮和基质细胞,观察细胞贴壁生长情况,同时对生长良好的原代细胞进行消化传代,进一步观察传代细胞的形态和生长情况。结果:角膜上皮、内皮和基质细胞均在培养4 8～72h贴壁生长,培养6～9d能形成良好单层,细胞形态典型,进行多次传代后细胞仍维持原有形态和功能,获得的细胞能进行长期培养。结论:为兔和牛角膜上皮、内皮和基质细胞体外培养。

Aim: To research the characteristics of corneal epithelial cells, endothelial cells and keratocytes of rabbit and bovine which were cultured in vitro, and to lay the foundation for tissue engineering of cornea. Methods: Rabbit and bovine corneal epithelial cells, endothelial cells and keratocytes were cultured by using digestion methods and the cells were subcultured after they became confluent. The growing characteristics were observed every day. Results: The epithelial cells, endothelial cells and keratocytes adhered and grew within 48～72 h and formed a confluent monolayer in 6～9 d. The shape of the primary isolated cells was typical, and the subcultural cells still maintained the characteristics of the primary cells. The cells grew very well and could be cultured for a long term. Conclusion: The research established a simple, highly efficient and economical method for in vitro culture of rabbit and bovine corneal epithelial cells, endothelial cells and keratocytes.