同一猴、兔眼角膜细胞成分原代培养的实验研究

目的建立从同一角膜片上分离培养角膜上皮、基质和内皮细胞的方法，为研究角膜细胞间的相互作用机制奠定基础。方法采用消化法分离培养猴眼角膜内皮细胞、上皮细胞和兔眼角膜上皮细胞．组织块培养法培养基质成纤维细胞和兔角膜内皮细胞。细胞接种于12孔培养板，并于培养不同时间行Wright染色检查细胞生长情况。结果采用消化法和组织块培养法能成功地培养猴、兔角膜上皮、基质和内皮细胞。猴、兔角膜内皮细胞培养1wk后，均能形成内皮细胞单层，细胞类似天然的六角形态．且以猴内皮细胞明显；猴角膜上皮细胞培养3～4d生长旺盛、但难以形成细胞单层，兔上皮细胞生长旺盛，1wk后已达融合状态，细胞呈膜状伸出板层伪足；猴、兔基质成纤维细胞易培养，1wk后已融合成单层细胞，细胞排列整齐，类似纤维走行样外观。结论从同一角膜材料中能分别培养出角膜3种细胞成份，消化法和组织块培养法相结合既能节省材料，又简单易行。

Laboratory study of the cornea cell primary culture from the same corneas of monkeys and rabbits

To establish the culture technique for comeal epithelial cells. kerato-cytes and endothelial cells from the same comeal material,and to lay a foundation for study ing the interaction mechanism among cornea cells. Methods The comeal endothelial cells.epithelial cells for monkeys and comeal epithelial cells for rabbits were cultured by using di-gestion methods. The keratocytes and endothelial cells for rabbits were cultured by tissue cul ture methods. All cells and small tissues were planted in 12-well culture plate and the culture cells morphological characteristics were observed by Wright staining during the various time periods. Results The comeal epithelial cells,keratocytes and endothelial cells from the same cornea could all be cultured successfully. In one week after the culture, the endothelial cells for monkeys and rabbits formed an adherent and conflunet monolayer, and the hexagon cells were the same as in vivo. The epithelial cells for monkeys grew very well at the 3rd or 4th day culture, but it did not form confluent monolayer. The epithelial cells for rabbits could form a confluent monolayer and the cells possessed a flattened lamellipodia. The keratocytes for monkeys and rabbits could be cultured easily and formed a monolayer and had a fibriform arrangement in one week after the culture. Conclusion Three kinds of cells from the same cornea can all be cultured successfully. The culture technique is practicable and simple.