新生大鼠视网膜神经干细胞的分离培养与诱导分化

CULTIVATION AND DIFFERENTIATION OF RETINAL NEURAL STEM CELLS IN NEONATAL RAT

目的 初步探讨新生大鼠视网膜神经干细胞的诱导分化。 方法 从新生大鼠视网膜分离具有自我更新能力的细胞克隆 ,传代。用脑源性神经营养因子 (BDNF)与血清单独或联合培养诱导其分化 ,采用免疫细胞化学及免疫荧光化学技术对分化前后的细胞进行鉴定。 结果 分离获得的细胞具有连续克隆的能力 ,表达nestin。分化后细胞分别表达多种神经元及神经胶质细胞的特异性标志物 ;BDNF与血清联合作用可使细胞向神经元分化的比例提高。 结论 新生大鼠视网膜内存在具有自我更新与多分化潜能特性的神经干细胞 ,BDNF作为辅助诱导剂 ,可促进其存活、分化、成熟 ,并可能诱导其向神经元方向分化。

Objective To isolate the retinal neural stem cells(RNSC) from the neonatal rat eye retina, and to study the induction and differentiation of the cells. Method Cell colonies with ability of proliferation were isolated from the retina of neonatal rat. Then brain derived neurotrophic factor(BDNF) or/and serum were used to induce them to differentiate. Immunofluorescence and immunocytochemistry with specific antibodies were used to identify undifferentied cells and differentied cells. Results Cell colonies showed ability of proliferation, and were able to differentiate into neurons and glia. Combination of BDNF with serum increased the ratio of neuron in differentiated cells.Conclusion There are retinal neural stem cells in neonatal rats. BDNF as a kind of assistant inducer, may help RNSC to survive, differentiate, mature, and probably induce them to differentiate into neurons.