大鼠视网膜Mller细胞受刺激后具有神经干细胞的特性

目的:研究并验证成年哺乳动物视网膜中Mller细胞经刺激后可以产生神经干细胞特性。方法:对6周龄VC大鼠右眼玻璃体腔内注射神经毒素混合液(含NMDA,kainate,FGF2和insulin)刺激Mller细胞,左眼玻璃体内注射PBS作为对照眼。1wk后取眼并分离Mller细胞体外原代纯化培养7d。流式细胞技术鉴定Mller细胞纯度,通过细胞免疫荧光组织化学和流式细胞技术鉴定神经干细胞标志物nestin在Mller细胞上的表达。结果:流式细胞分析结果显示,本实验分离培养的原代Mller细胞纯度可达96%以上;细胞免疫荧光组织化学显示激活后Mller细胞能表达nestin,流式细胞技术证明,激活后培养的细胞中表达nestin的细胞可达总细胞量的53.5%。结论:成年大鼠视网膜Mller细胞经神经毒素在体刺激后可以产生神经干细胞特性。

Research on neural stem cell properties of Müller cells in rat

AIM:To investigate and testify that the neural stem cell properties of Müller cells can be activated in adult mammal. ·METHODS: Introvitreal injected mixed solution of neurotoxins(including NMDA, Kainate,FGF2 and insulin) to 6 weeks old VC rat to activate Müller cells.One week later,the activated Müller cells were gathered,cultured and then purified. The FACS analysis was carried out to assess the purity of the cultured Müller cells and the immunocytochemical analysis and the FACS analysis were used to find out neural stem cell marks on Müller cells. ·RESULTS: The purity of the cultured Müller cells was above 96% and more than 53.5% of all cells were nestin positive. ·CONCLUSION: The neural stem cell properties of retinal Müller cells can be activated in adult rat.