不同浓度重组人促红细胞生成素对神经干细胞体外培养增殖的影响

背景:重组人促红细胞生成素是一种糖蛋白,近年的研究表明其对神经细胞的许多功能活动均具有调节作用.目的:观察不同浓度重组人促红细胞生成素对神经干细胞体外培养增殖的影响.方法:提取新生SD大鼠神经干细胞,用含不同浓度(5,50,500 U/mL)重组人促红细胞生成素和20 μg/L碱性成纤维细胞生长因子的无血清培养基进行培养,以不含重组人促红细胞生成素无血清培养基为对照组.细胞培养7 d后计算神经干细胞克隆形成率,培养10 d后计数NSE和GFAP免疫阳性细胞数.结果与结论:添加重组人促红细胞生成素组细胞增殖较快,最终神经球的数量多于对照组,以50 U/mL重组人促红细胞生成素组作用显著;50 U/mL重组人促红细胞生成素组的生长速度显著快于对照组.50 U/mL重组人促红细胞生成素组中NSE和GFAP免疫阳性细胞明显多于对照组(P<0.01).结果表明重组人促红细胞生成素对神经干细胞体外培养增殖有促进作用,尤以适中浓度(50 U/mL) 作用更加明显.

Effects of different concentrations of recombinant human erythropoietin on proliferation of neural stem cells cultured in vitro

BACKGROUND: Recombinant human erythropoietin (rhEPO) is a glycoprotein. Recent studies have demonstrated that rhEPO regulates many functional activities of neural cells. OBJECTIVE: To investigate the effects of different concentrations of rhEPO on proliferation of neural stem cells (NSCs) cultured in vitro. METHODS: Newborn Sprague-Dawley rat NSCs were harvested and cultured with serum-free culture medium containing different concentrations (5, 50, 500 U/mL) of rhEPO and 20 μg/L basic fibroblast growth factors (5, 50, and 500 U/mL rhEPO groups) and serum-free culture medium only containing 20 μg/L basic fibroblast growth factors (control group). After 7 days of culture, the cloning efficiency of NSCs was calculated. After 10 days of culture, neuron specific enolase (NSE)-and glial fibrillary acidic protein (GFAP)-immunoreactive cells were quantified. RESULTS AND CONCLUSION: In the rhEPO groups, cells proliferated rapidly, and the number of NSC microspheres was greater, in particular in the 50 U/mL rhEPO group, compared with the control group. NSCs grew faster in the 50 U/mL rhEPO group than in the control group. The number of NSE-and GFAP-immunoreactive cells was greater in the 50 U/mL rhEPO group than in the control group (P<0.01). These findings suggest that rhEPO promotes the in vitro culture and proliferation of NSCs, in particular 50 U/mL rhEPO.