Matrigel胶对大鼠海马神经干细胞增殖及分化的影响

Effect of Matrigel on the proliferation and differentiation of rat hippocampal neural stem cells

目的:观察Matrigel胶对大鼠海马神经干细胞(NSCs)增殖和分化的影响。方法:自出生24 h内SD大鼠海马组织提取NSCs,取培养第3代细胞,通过Nestin蛋白免疫细胞化学染色进行鉴定。取第3代细胞分为4组,A组接种于多聚赖氨酸包被24孔板,加入正常培养基;B组接种于多聚赖氨酸包被24孔板,加入含胎牛血清的DMEM/F12培养基;C组接种于Matrigel胶内,加入正常培养基;D组接种于Matrigel胶内,加入含胎牛血清的DMEM/F12培养基。培养1周后,通过免疫荧光染色方法检测NSCs中巢蛋白(Nestin)、β-tubulinⅢ、胶质纤维酸性蛋白(GFAP)的表达情况。结果:第3代培养细胞Nestin蛋白阳性,证实为NSCs。A组NSCs分化缓慢,B、C、D组NSCs分化迅速,C、D两组中NSCs形成立体的网状结构。4组NSCs中Nestin、β-tubulinⅢ、GFAP蛋白免疫荧光染色阳性率差异有统计学意义(F培养基分别为3 918.000、437.023和3 395.000,F包被分别为291.234、4 833.000和50.081,P均<0.001),表明B、C、D组NSCs分化显著,且C、D两组中NSCs向神经元分化趋势明显,C组NSCs向神经元分化趋势最强。结论:Matrigel胶具有诱导NSCs向神经元分化的作用。

Aim：To observe the effect of Matrigel on the proliferation and differentiation of rat hippocampus neural stem cells（NSCs）.Methods：NSCs were extracted from newborn SD rats′hippocampus, and the cells after 3 generations were identified through Nestin cytochemical staining .The NSCs were allocated into four groups：group A was inoculated on 24-well plate coated by poly-lysine and added normal culture medium;group B was inoculated on 24-well plate coated by poly-lysine and added DMEM/F12 medium containing fetal bovine serum; group C was inoculated on 24-well plate coated by Matrigel and added normal culture medium;group D was inoculated 24-well plate coated by Matrigel and add-ed DMEM/F12 medium contarning fetal bovine serum .After a week, the expressions of Nestin ,β-tubulinⅢand glial fibril-lary acidic protein（GFAP） by immunofluorescence staining.Results：Nestin protein was positive in the cells after 3 genera-tions, which confirmed that they were NSCs .NSCs in group A differentiated slowly , while group B,C and D differentiated rapidly.Especially, NSCs in group C and D formed three-dimensional network structure .The immunofluorescence staining positive rate of Nestin,β-tubulinⅢ and GFAP among the four groups had statistical significance （ Fmedium =3 918.000, 437.023, and 3 395.000 respectively,Fcoat =291.234,4 833.000, and 50.081 respectively,P〈0.001）,which showed that NSCs in group B,C and D were significantly differentiated , and the trend of NSCs differentiated into neurons was more obvi-ous in group C,D,especially group C.Conclusion：Matrigel could induce NSCs to differentiate into neurons .