兔MSCs移植对兔角膜缘干细胞形态、增殖、分化的影响  

Effect of MSCs transplantation on morphology,proliferation,and differentiation of rabbit corneal limbal stem cell

作　　者：余锦强[1] 李凌[1] 李儒华[1] 柯峰[1] 李芳[1] 邵杰[1] 

机构地区：[1]湖北医药学院附属人民医院眼科,湖北十堰442000

出　　处：《西南国防医药》2015年第2期134-137,共4页Medical Journal of National Defending Forces in Southwest China

基　　金：十堰市科学技术研究与开发项目[十科发(2010)23号2010-014z]

摘　　要：目的探讨兔骨髓间充质干细胞(MSCs)移植对兔角膜缘干细胞形态、增殖、分化的影响。方法无菌条件下取兔骨髓,分离得到兔MSCs。取培养3代后MSCs消化后制成细胞悬液,接种于羊膜植片上,常规培养。建立兔角膜碱烧伤干细胞缺损模型4 w后,将18只新西兰兔随机分为空白组、MSCs移植组和模型组,每组6只。MSCs移植组和模型组分别用尼龙缝线将空白和带有兔MSCs的羊膜基质植片间断缝合固定于浅层巩膜上,空白组不做处理。术前、术后观察实验兔眼表形态,并进行评分。术后4 w处死动物,免疫组化检查细胞角蛋白AE5表达情况。结果筛选得到的兔MSCs细胞均一性达98.12%;术后MSCs移植组兔眼表形态综合评分显著降低,而模型组无显著变化,同期与模型组综合评分相比,均显著低于模型组(P<0.05);免疫组化检测结果显示,空白组和MSCs移植组AE5阳性表达率均为100.00%,显著高于模型组(P<0.05)。结论以羊膜基质为载体移植MSCs治疗角膜缘干细胞缺乏症,可增殖分化为角膜上皮细胞而发挥功能,疗效显著,可为其临床应用提供理论基础。Objective To explore the effects of rabbit bone marrow mesenchymal stem cells （MSCs）transplantation on morphology, proliferation, and differentiation of rabbit corneal limbal stem cell. Methods Rabbit bone marrow was obtained under sterile conditions and MSCs were obtained by isolation. Cell suspension was prepared after digestion of MSCs cultured for three generations and inoculated in the amniotic membrane graft, and conventional culture was provided. And then, a rabbit corneal alkali burn stem cell defect model was built. After four weeks, 18 New Zealand rabbits were randomly divided into blank group, MSCs transplantation group and model group（ n = 6 per group）. For the M SCs transplantation group and model group, nylon suture was used to fix the amniotic membrane grafts with and without MSCs discontinuously to the superficial sclera, and no treatment was provided for the blank group. The preoperative and postoperative ocular surface morphology of the experimental rabbits were observed and scores were kept. The rabbits were put to death after four weeks since the operation, and immunohistochemistry method was used to examine the expression of cytokeratin AE5. Results The homogeneity of rabbit MSCs obtained by screening reached to 98. 12%; The comprehensive scores of postoperative ocular surface morphology of the rabbits in the MSCs transplantation group decreased significantly, while the model group did not change significantly;compared with the model group, the comprehensive scores in the same period were all significantly lower than that in the model group （ P 〈 0.05 ） ; immunohistochemistry results showed that the positive expression rates of AE5 in the blank group and MSC transplantation group were 100.00%, significantly higher than that in the model group（ P 〈 0.05 ）. Conclusion Taking amniotie membrane matrix as a cartier for MSCs transplantation in treatment of corneal limbal stem cell deficiency is available for proliferation and differentiation of corneal epithelial cells, and the

关 键 词：骨髓 间充质干细胞 角膜缘 干细胞缺损 增殖 分化 

分 类 号：R779.65[医药卫生—眼科]