摘要
目的:观察人骨髓间质干细胞(MSCs)移植到兔角膜基质后的分化发育情况,探讨MSCs分化为角膜上皮细胞的可行性。方法:24只新西兰兔随机分为2组。实验组:将人MSCs接种在保存人羊膜上培养4d,用5-溴脱氧尿嘧啶(BrdU)标记后移植到兔角膜基质;对照组:采用保存羊膜移植到兔角膜基质。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,检查移植到兔角膜基质的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角膜基质的带有BrdU标记的细胞角蛋白K3/12(CK3/CK12)和角蛋白K13(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。羊膜负载MSCs移植到兔角膜基质表面,术后免疫组织化学检测角膜上皮层CK3/CK12表达阳性,CK13表达阴性,在重建的角膜上皮层可检测到BrdU核阳性细胞并同时表达角膜上皮细胞特异性表面标志蛋白K3/K12,未发生免疫排斥反应,未见异常增殖细胞。结论:羊膜负载MSCs移植到兔眼表角膜基质后,MSCs能存活、增殖并向角膜上皮样细胞分化。
AIM： To observe the differentiation and development of human mesenchymal stem cells （MSCs） transplanted onto corneal stroma of rabbits and investigate the feasibility of MSCs differentiated into corneal epithelium like cells. METHODS： 24 New Zealand albino rabbits were randomly divided into 2 groups ： The human MSCs combined with amniotic membrane were transplanted onto the experimental animals, and the controls were transplanted with the amniotic membrane only. The MSCs were cultured on preserved human amniotic membrane for 4 days and labeled with 5＇ - Bromo - 2＇-deoxyuridine （BrdU）, then the cells were transplanted onto the surface of the corneal stroma of the 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： When the MSCs cultured on amniotic membrane were transplanted onto the surface of the corneal stroma of rabbits, the corneal epithelium were positive in CK3/CK12 staining and negative in CK 13 staining, revealed by immunohistochemical examination. The BrdU positive cells in the reconstructive corneal epithelium were found and showed positive in CK3/CK12 staining. CONCLUSION： After transplanted onto the corneal stroma of rabbits with human amniotic membrane, the MSCs survive, proliferate and differentiate into corneal epithelium like cells.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2007年第5期999-1003,共5页
Chinese Journal of Pathophysiology
基金
广东省自然科学基金资助项目(No031734)
关键词
骨髓间质干细胞
羊膜
上皮
角膜
细胞分化
Bone marrow mesenchymal stem cells
Amnion
Epithelium, corneal
Cell differentiation
