视网膜祖细胞干细胞特性及移植入视网膜后的研究

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-Di(?) 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。

Study of retinal progenitor cell characteristics and its transplantation into vitreous cavity

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 degen- eration 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-DiI labeling and retinal progenitors were transplanted into vitreous cavity.Re- sults:Retinal progenitors expressed nestin,Flk-1,Pax6 and Notchl mRNA and incorporated BrdU.Retinal pro- genitors differentiated and expressed retinal cells specific proteins.After transplantation,retinal progenitors sur- vived,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.