增龄对角膜基质载体培养的角膜缘干细胞生物学特性的影响

目的 观察增龄对新西兰兔角膜缘干细胞在同种异体角膜基质上生长、增殖的影响.方法 用酶消化法将不同年龄新西兰兔角膜缘组织制成细胞悬液培养,以相同的细胞密度种植于不同年龄的新西兰兔的角膜基质上,观察角膜缘干细胞在角膜幕质上的生长情况.原位杂交检测ABCG2、ANp63的表达,IPP5.1软件分析图像,流式细胞术测细胞周期,运用SPSS13.0软件对析因设计资料进行方差分析.结果 角膜缘干细胞供体年龄各水平差异有统汁学意义,相同时间内角膜基质培养的青年兔角膜缘干细胞在角膜基质单位面积卜的牛长总而积均较中年和老年高,增殖期细胞所占比例大;而角膜基质供体年龄各水平差异与角膜缘干细胞和角膜基质的交互作用均无统计学意义.结论 在同种异体角膜基质载体上培养的角膜缘干细胞的增殖能力随着年龄的增加而降低.

Effects of aging on the biological characteristics of cultured limbal stem cells on the corneal stroma as a carrier in vitro

Objective To investigate the effects of aging on the growth and proliferation of cultured limbai stem cells(LSCs)from New Zealand rabbits in aliogeneic corneal stroma.Methods Limbal epithelium harvested from New Zealand rabbits of different ages were enzymatically cleft into single cell suspension and cultured,and limbal stem cells were seeded at a same cell density in the corneal stroma of different age of New Zealand white rabbits.The growth status of LSCs seeded in the corneal stroma was observed while identification was performed using in situ hybridization,image was analyzed in IPP5.1 software,cell cycle was examined by flow cytometry,and the experimental data was analysed by ANOVA of factorial design.Results The difference in the level of LSCs donor age has statistical significance,as the total growth area of LSCs in unit area of corneal stroma and the ratio of proliferating cells were the highest in the young rabbits compared with the middle-aged and elder rabbits;the difference in the level of corneal stroma donor age and the interation of LSCs and corneal stroma have no statistical significance.Conclusion The proliferation capacity of limbal stem cells seeded in allogeneic corneal stroma was shown to decrease with aging.