兔结膜基质诱导人骨髓间质干细胞分化的初步研究

 【目的】观察人骨髓间充质干细胞（mesenchymal stem cells，MSCs）在保存人羊膜及兔眼表结膜基质上分化的情况。【方法】24只新西兰兔随机分为2组。实验组：培养有人MSCs的羊膜移植组，将人MSCs接种在保存人羊膜上培养4d，用5-溴脱氧尿嘧啶（BrdU）标记后移植到兔眼表结膜缺损区；对照组，单纯羊膜移植组。分别于移植后1，2，3，4，6和8周，摘取各组实验眼行组织学和免疫组织化学检查，组织学检查移植到兔结膜基质的MSCs的存活、形态变化以及移植局部的反应等情况；免疫组织化学检测移植到结膜基质的带有BrdU标记的细胞角蛋白3／12（CK3／CKl2）和角蛋白13（CK13）的表达。【结果】MSCs接种到羊膜后能在羊膜上生长，与羊膜共培养4d后，MSCs贴附羊膜生长迅速，组织学特征无明显改变。用羊膜负载MSCs移植到兔结膜缺损区，术后免疫组织化学检测结膜上皮层CK13表达阳性，CK3／CK12表达阴性，在重建的结膜上皮层可检测到BrdU核阳性细胞，未见异常增殖细胞。【结论】人羊膜可作为MSCs的载体，采用人羊膜负载MSCs行兔结膜移植，MSCs能在兔结膜上皮层存活并增殖。

Preliminary Study on Differentiation of Human Mesenchymal Stem Cells Induced by Conjunctival Stroma of Rabbits

Objective] To investigate the differentiation of human mesenchymal stem cells(MSCs)induced by human amniotic membrane and conjunctival stroma of rabbits.Methods] Twenty-four New Zealand albino rabbits were equally and randomly divided into 2 groups:the experimental group was the human MSCs combined with amniotic membrane transplantation group and the control group was the amniotic membrane transplantation group.The MSCs were cultured on preserved human amniotic membrane for 4 days and labeled with 5'-Bromo-2'-deoxyuridine(BrdU),then they were transplanted on the scleral area of New Zealand albino rabbits.The eyeballs were taken off after 1,2,3,4,6,and 8 weeks.The growth and differentiation of human MSCs were observed by histopathological and immunohistochemical examination.Results] The MSCs grew well on amniotic membrane.The histological characters of MSCs did not changed.When the MSCs cultured on amniotic membrane were transplanted on the scleral area of rabbits,Immunohistochemical examination revealed that the conjunctival epithelial were positive in cytokeratin 13 staining and negative in CK3/CK12 staining.The BrdU positive cells in reconstructive conjunctival epithelial were found.There were no abnormal proliferative cells.Conclusions] The MSCs may be cultured on human amniotic membrane in vitro.When they were cultured on amniotic membrane and transplanted onto the scleral area of rabbits they could survive,proliferate without abnormal proliferation.