培养兔自体角膜缘干细胞移植的实验研究

目的 观察以羊膜为载体的兔角膜缘于细胞膜片移植治疗兔角膜缘于细胞缺损的效果。方法 制造兔角膜缘干细胞缺损的动物模型，以右眼为实验眼，从兔左眼取角膜缘组织，将兔角膜缘干细胞消化下来，接种于铺有羊膜的无菌六孔培养板中，待细胞形成多层角膜上皮细胞后，对兔角膜缘干细胞缺损动物模型行角膜缘干细胞羊膜移植术，并对治疗后的角膜进行裂隙灯及病理学检查。结果 临床和组织病理学染色证实：体外培养的兔角膜缘干细胞可在羊膜上继续增生、分化为多层角膜上皮细胞；角膜缘干细胞移植术后兔角膜缘轻度充血、角膜上皮完整、基质细胞浸润减轻、新生血管减少或消失。结论 应用角膜缘干细胞羊膜移植术可恢复其角膜上皮结构的完整性，减少角膜新生血管的形成，维持角膜缘的细胞屏障功能。

An experimental study on autograft transplantation with cultured limbal stem cells on amniotic membrane

Objective To study the treatment of rabbit limbal stem cell deficiency by cultured limbal stem cells growing on amniotic membrane for autograft transplantation. Methods To make rabbit models with rabbit limbal stem cells deficiency which right eyes were experimental ones. After the the rabbit corneal stem cells were digested, they were planted on amniotic membrane. When they differentiated and formed multiple cell layers on amniotic membrane. Corneal limbalstem cells and amniotic membrane were tramsplanted on the limbal and scleral area of the rabbit model with rabbit limbal stem cell deficiency. The corneal changes were observed by a slitlamp everyday, and the corneal pathological changes were examined. Results clinical and pathological examination verified that cultured rabbit corneal stem cells continued to proliferate, differentiate and form multiple cell layers on amniotic membrane. After transplantation with cultured stem cells and amniotic membrane, the rabbit epithelium showed corneal phenotype and progressive decrease of vascularity and stromal infiltration in the limbal and peripheral zone. Conclusion Autograft transplantation with cultured limbal stem cells can restore corneal limbal stem cell composition, decrease neovascularization, maintain limbal cellular barrier function.