兔视网膜色素上皮细胞移植的实验研究

Study of transplantation of retinal pigment epithelial cells

目的 用改良的二步酶法制备视网膜色素上皮 (RPE)细胞悬液 ,不做玻璃体切割 ,直接进行视网膜下间隙的移植。方法 用酶Ⅰ (含 2 2 0kU/L透明质酸酶和 0 .1 %胰蛋白酶 )分离视网膜神经上皮层与RPE之间的联系 ,再用酶Ⅱ (含 0 .2 5 %胰蛋白酶 )将RPE从玻璃膜上分离下来获取悬浮的RPE细胞 ,用特制的移植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。电镜观察术后 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膜形成镶嵌关系。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

Objective To explore the preparation of retinal pigment epithelial(RPE)cells via the improved two step enzymatically digestive way and transplantation of RPE cells into subretinal space through a modified internal approach excluding vitrectomy. Methods The neural retina was isolated from RPE by enzyme Ⅰ containing 220 kU/L hyaluronidase and 0.1% trypsin, then the RPE cells were isolated from the Bruch's membrane by enzyme Ⅱ containing 0.25% trypsin, and after that the resembled RPE cells were injected into the host's subretinal space with a special equipment after producing an artificial bleb detachment of neural retina. Results Two weeks after transplantation surgery, the transplanted RPE cells survived as a quasi monolayer under the host's subretinal space and there was no obvious inflammation. Seven weeks after transplantation surgery, the electron microscopy showed the transplanted REP cells were attached to the host's Bruch's membrane and survived in contiguity with healthy host's outer segment. Conclusion The transplanted RPE cells can be transplanted into the host's subretinal space and survive at least for seven weeks through the modified internal approach.