兔視網膜色素上皮细胞移植的實驗研究

目的 探索用改良的二步法酶法制备视网膜色素上皮(RPE)细胞悬液,经改良的内路法,不经玻璃体切割的情况下进行视网膜下间隙的移植 方法 用酶Ⅰ(含220kU/L透明质酸酶和0.1％的胰蛋白酶)松解视网膜神经上皮细胞与RPE细胞之间的联接,再用酶Ⅱ(含0.25％胰蛋酶)从Bruch膜上离散RPE细胞,制成细胞悬液 用特制的注射针头,通过人为造成的局限性视网膜脱离区将其移植入受体视网膜下间隙 结果 术后2周供体RPE细胞层呈单层排列于受体视网膜下间隙,并存活,未见明显炎症反应 电镜观察,术後7周供体RPE细胞位于受体Bruch膜上并与受体感光细胞外节盘模型成镶嵌关系结论改良的内路法可以将供体RPE细胞成功移植在受体眼视网膜下间隙,并至少存活7周,改良的内路法渴望为视网膜移植的实验室研究提供一条可供借鉴的途径。

An experimental study of transplantation of rabbit retinal pigment epithelial cells

Objective To study preparation of retinal pigment epithelial (RPK)cells via the improved lwo-step enzymateally digestive way and transplantation of RPK fells into subretinal space through a modified internal approach without vireetomy. Methods Enzyme (containing 220kU/L hyaluromdase and 0.1% trypsin) was used to loose the connection between the RPE cells and the neural retina. Enzyme (containing 0.25% trypsin) was used to dissociate RPE cells from Bruch's membrane. Then the resembled RPE cells was injected into the host's subretinal space with a special equipment after an artificial bleb detachment of neural retina was produced. Results Two weeks after transplantation surgery, the transplanted RPK cells survived as a quasi-monolayer in the host's subretinal space and obvious inflammation didn't occur. Seven weeks after transplantation surgery, the the electron microscopy showed the transplanted RPK cells attached to the host's Bruch's membrane and surived in contiguity with health) host's outer segment. Conclusion The transplanted RPK cells can be transplanted into the host's subretinal space and survive at least for seven weeks through the modified internal approach. The modified internal approach promises an advisable method for the laboratory research of RPE transplantation.