腺苷促进体外培养神经干细胞增殖作用研究

Adenosine Promotes Proliferation of Neural Stem Cells in vitro

目的研究腺苷促进体外培养神经干细胞(NSCs)增殖作用。方法取新生小鼠大脑进行神经干细胞原代培养,观察神经球生成情况,特异性蛋白质免疫细胞化学染色和BrdU标记鉴定并判断其增殖能力;将腺苷按0.4,2.0,10.0μmol.L-13个浓度加入神经干细胞培养基,同时设立溶媒对照组和阳性对照组,通过观察神经球形态、神经球生成数目及MTT法定量比较,分析不同浓度腺苷对神经干细胞分裂增殖的影响。结果培养物中有大量神经球生成,神经干细胞特异性蛋白质免疫细胞化学染色呈阳性,BrdU标记亦呈阳性反应,所培养的细胞为神经干细胞,具有增殖能力;腺苷中、高浓度组神经球数目、MTT比色结果明显高于溶媒对照组。结论腺苷具有促进NSCs增殖作用。

Objective To study the effects of adenosine on proliferation of neural stem cells in vitro. Methods Primary neural stem cells （ NSCs ） from newborn mice were cultured, neurospheres were observed, and immunocytochemical staining of characteristic protein and BrdU labeling were performed to identify NSCs and their proliferative properties. Adenosines with 0.4, 2.0, 10.0 μmol · L^-1 were respectively added to the cultured NSCs, and at the same time, a vehicle control and positive control were set up. The morphology of NSCs was observed with the convert phase microscope, the neurosphere numbers were counted, and NSCs viability was determined with MTT assay in order to analyze the proliferation of NSCs by adenosine in vitro, Results A great deal of neurospheres were formed in cultured cells, the characteristic protein of NSCs and BrdU labeling were positive, which suggesting that the cultured cells were NSCs, and had the potential of self-renewal and self- proliferation. The number of neurosphere and the A values of 2.0 μmol · L^-1 and 10. 0 μmol · L^-1 adenosine treated group, respectively were significantly higher than those of the vehicle control group. Conclusion Adenosine shows effects of promoting the proliferation of neural stem cells.