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目的 探讨离体培养的角膜干细胞增殖分化特性 ,观察干细胞羊膜移植片治疗角膜缘功能障碍的效果。方法 用克隆形成率 (colony formingefficiency ,CFE)、免疫组织化学染色等方法观察培养角膜干细胞的增殖力和分化表达状况。角膜干细胞培养后接种生长于羊膜上进行干细胞羊膜片移植 ,观察角膜上皮、组织浸润和新生血管情况。结果 原代和传第 1代角膜干细胞具有高增殖力 ,部分表达角质蛋白K3,角膜干细胞在羊膜上能够继续增殖分化为多层细胞结构的角膜上皮细胞层 ;角膜干细胞移植术后可以在角膜比存活 ,角膜比移植片紧密贴附 ,角膜上皮完整 ,经度基质炎性浸润 ,浅层新生血管减少 ,深层新生血管充盈减轻 ,睑球粘连得到松解。结论 角膜缘干细胞羊膜片移植治疗角膜缘功能障碍能够使患者获得进一步进行角膜移植的条件。
Objective To investigate the proliferation and differentiation characteristics of cultured corneal stem cells in vitro and observe the effect of stem cell amniotic membrane transplantation for the treatment of limbal dysfunction. Methods Colony formingefficiency (CFE) and immunohistochemical staining were used to observe the proliferation and differentiation of cultured keratocytes. After culture of corneal stem cells, the cells were inoculated on the amniotic membrane for transplantation of stem cell amniotic membrane and the corneal epithelium, tissue infiltration and neovascularization were observed. Results Primary and passage 1 keratocytes had high proliferative capacity and partially expressed keratin K3. Corneal stem cells could continue to proliferate and differentiate into multi-layer corneal epithelial cells in amniotic membrane. After corneal stem cell transplantation, Survival, the cornea is closely attached to the graft, the corneal epithelium is intact, the infiltration of longitude stroma, the decrease of superficial neovascularization, the reduction of deep neovascularization filling and the adhesion of the symblepharon are released. Conclusion The limbal stem cell transplantation of amniotic membrane for corneal limbal dysfunction can make the patients obtain the condition of further corneal transplantation.