摘要
目的 评价深低温冷冻保存兔眼角膜缘干细胞活性。方法 12只 (12眼 )新西兰白兔 ,制备 2mm周边角膜和 2mm球结膜的环形浅层角膜缘植片及角膜干细胞缺乏眼表疾病模型。角膜缘植片通过程序降温仪程序降温后 - 196℃深低温保存 ,30天和 6 0天后行角膜缘自体移植。组化及免疫组化证实深低温保存的角膜缘干细胞活性。结果 角膜干细胞缺乏导致角膜上皮愈合延迟 ,上皮结膜化 ,PAS染色阳性 ,AE5染色弱阳性 ;基质水肿 ,角膜血管化。深低温保存角膜缘干细胞自体移植能促进角膜上皮愈合 ,为角膜表型 ,PAS染色阴性 ,AE5染色阳性。基质水肿逐渐吸收 ,新生血管消退、变细。结论 深低温保存法能保存角膜缘干细胞活性 ;能随时按需为临床提供活性角膜缘材料 ;为深低温保存角膜缘干细胞移植治疗临床眼表疾病提供了实验依据。
Objective:To evaluate the viability of limbal stem cells with cryopreservation.Methods:Limbal grats include 2mm of peripheralcornea and 2mm of conjunctiva and ocular surface disorders(OSDs) models of corneal stem cells deficiency were created in right eyes of 12 rabbits. The grafts were cryopreserved in -196℃ liquid nitrogen by program controlled Bio Freezer. The limbal autograft transplantations for ocular surfacereconstruction were performmed after cryopreserved 60 days and 30 days. The viability of limbal stem cells with cryopreservation was proved by histochemistry and immunohistochemistry.Results:The stem cells deficiency results in corneal epithelium delay healing, it showed conjunctival epithelial phenotype by a positive staining PAS and a weak positive staining of the immunohistochemistry of monoclonal antobody AE5 and the stromatic swelling and neovascularization. The cryopreserved limbal autograft transplantion can promote corneal epithelium healing, it showed corneal epithelial phenotype by a negative staining PAS and a positive staining of the immunohistochemistry of monoclonal antobody AE5. It was observed that corneal stromal swelling and neovascularization were markedly decreased.Conclusion:Cryopreservation in liquid nitrogen can preserve the viability of limbal stem cells, and provide viable limbal grafts to treat OSDs at any time. It provide a experimental evidence for cryopreserved limbal stem cells to treatment OSDs.
出处
《重庆医学》
CAS
CSCD
2000年第6期481-482,共2页
Chongqing medicine
基金
美国中华医学基金会(CMB)资助!(基金号 :90 543)
