目的:研究视网膜祖细胞的干细胞特性及移植入视网膜后的存活和迁移。方法:体外培养胎龄18d大鼠的视网膜细胞,用RT-PCR、细胞免疫荧光方法鉴定其增殖分化;成年SD大鼠腹腔注射N-甲基-N-亚硝基脲形成视网膜感光细胞退化的动物模型,培养的视网膜祖细胞用CM-DiI标记后,移植入模型鼠的玻璃体腔。结果:视网膜祖细胞体外表达中间神经丝蛋白nestin;表达Flk-1、Pax6及Notchl的mRNA;能掺入BrdU;分化后表达视网膜各类细胞特异性蛋白;移植后在实验组大鼠视网膜能存活及迁移,而在对照组中仅聚集在玻璃体腔。结论:视网膜祖细胞具有干细胞特性,移植入受损伤视网膜后,能存活、整合及迁移。
Objective: To investigate stem cell characteristics of retinal progenitors and to observe their survival and migration after transplanted into vitreous cavity. Methods: E18 fetus rat retinal cells were cultured in vitro; the proliferation and differentiation were identified by RT-PCR and cytoimmunochemistry; photoreceptor degeneration model was made by i. p injection N-methyl-N-nitrosourea at a dose of 60 mg/kg to adult SD rats. Retinal progenitors were traced by CM-Dil labeling and retinal progenitors were transplanted into vitreous cavity. Resuits: Retinal progenitors expressed nestin, Flk-1, Pax6 and Notchl mRNA and incorporated BrdU. Retinal progenitors differentiated and expressed retinal cells specific proteins. After transplantation, retinal progenitors survived, integrated and migrated into retinas of model animals, while they only aggregated in control. Conclusion: Retinal progenitors display some properties of stem cells; they can survive, integrate and migrate into damaged retina.