摘要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-forming efficiency, 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.

abstractsObjective To investigate the proliferation and differentiation of cultured corneal stem ce lls and determine the effect of corneal stem cells cultured on amniotic membrane s on the limbal area for treating corneal burns. Methods The proliferation and differentiation of corneal stem cells in vitro had been ex amined using colony-forming efficiency and immunohistochemistry. The stem cell s had been cultured on amniotic membranes and transplanted to the limbal area fo r treating corneal burns. Results Corneal stem cells had a high proliferation capacity in primary and first passag e, cytokeratin 3 was not expressed in primary culture but partly in first passag e. The stem cells could proliferate to form cell layer on an amniotic membrane . When transplanted, stem cells could survive on limbus. After transplantation , ocular inflammation resolved, the cornea re-epithelialized, the stromal opaci ty reduced, the superficial neovascularity was lessened and the conjunctival for nix re-established. Conclusions Ocular surface conditions could be improved by allograft of corneal stem cells c ultured on amniotic membranes.