兔角膜缘上皮细胞培养联合羊膜行自体移植的实验研究

目的：用体外培养的角膜缘上皮细胞联合人羊膜的方法，行自体移植治疗角膜缘干细胞完全缺乏的兔眼。方法：制作10只兔右眼于细胞缺乏的模型，取其中7只兔的左眼上方角膜缘取一小块组织进行体外培养，并传代在无上皮细胞的人羊膜上。1个月后，手术切除10只兔受伤眼角膜表面被覆的新生血管膜和结膜上皮，7眼接受了含自体培养上皮细胞的羊膜移植，另3眼仅移植无上皮细胞的羊膜。结果：1周后原代培养的角膜缘上皮细胞融合，传代至无上皮细胞的人羊膜上继续生长2～3周，角膜上皮细胞可牢固的附着于羊膜上。移植了含有角膜上皮细胞的羊膜的兔子，术后早期都形成了角膜上皮化，并明显抑制了新生血管的再生，而接受羊膜移植的兔子，术后又出现明显的新生血管。结论：利用无上皮细胞的羊膜作为载体，培养角膜缘上皮细胞并自体移植可以恢复角膜上皮化、抑制新生血管生长。

Experimental Study of Cultivated Rabbit Corneal Limbal Epithelial Cells Autograft With Amniotic Membrane Grafts

Objective: To study in vitro cultivated limbal epithelial cells autograft with amniotic membrane grafts for total limbal stem cell deficiency of rabbit eyes. Methods: Ten rabbit eye models of limbal stem cell deficiency were made. A small piece of tissue was obtained from upper limbal area of each left eye of seven models to be cultivated in vitro and passaged on acellular human amniotic membrane (AM) . A month later, surgical dissection was performed to remove the neovascular membrane and conjunctival epithelia on the corneal surface of the ten damaged eyes. Then seven eyes received cultivated epithelial cells autograft with amniotic membrane grafts and the other three only underwent acellular AM transplantation. Results: A week later, the primary cultivated limbal epithelial cells were fused and continued to grow for 2 to 3 weeks after being passaged on acellular human AM. The corneal epithelial cells were attatched to the AM very firmly. Corneal epithelialization and significant suppression of reneovascularization were observed on the rabbits which received cultivated corneal epithelial cells autograft with AM grafts at the early stage of the postoperative period, while significant reneovascularization was observed on the rabbits which underwent AM transplantation. Conclusion: Cultivated limbal epithelial cells autograft on acellular AM could restore epithelialization and suppress neovascularization.