兔角膜基质细胞的体外培养及生物学特性

目的改进兔角膜基质细胞培养技术,观察其生物学特性。方法　先同时消化角膜上皮及内皮层,然后刮去上皮、内皮、前弹力和后弹力层,接着将剩余的基质层剪碎消化、接种,采用MTT法观察细胞生长情况,并对比有、无血清培养基对细胞生长的影响。结果　角膜基质细胞10d后形成细胞单层,增殖旺盛。有血清培养基培养到第8天左右达到生长高峰,无血清培养基培养的细胞生长较慢,第9天左右达到生长高峰。结论　改进的角膜细胞培养技术简便、成功率高,培养出的角膜基质细胞生长良好。血清对角膜基质细胞生长有一定的影响。

In vitro culture and study of the biological characteristics of rabbit keratocytes

Objective To improve the technique for culturing rabbit keratocytes in vitro and investigate the biological characteristics of these cells. Methods Fresh rabbit corneas were obtained and the epithelial and endothelial cells were removed after digestion with trypsin. The stroma was rinsed, minced, and incubated in DMEM/F12 medium supplemented with 10% fetal bovine serum and the biological characteristics of the keratocytes were observed with MTT assay and compared with those of the cells in serum-free culture media. Result On about the third day of incubation, some keratocytes germinated from the stro-mal tissues and migrated onto the flask surface presenting fibroblast-like arrangement with spindle-shaped appearance. The keratocytes became confluent after 10 day's incubation and the peak of cell proliferation occurred on day 8 in the presence of serum in the media, while in the absence of serum, the peak took place on day 10. Conclusion The method improved for in vitro keratocyte culture is convenient and effective, and the presence of serum in the media may to some degree affect the growth of the keratocytes.