骨髓基质细胞和神经干细胞体外共培养的实验研究

The Experimental Study of Mesenchymal Stem Cells in Co-Culture with Neural Stem Cells in Vitro

目的:研究神经干细胞(NSCs)和骨髓基质细胞(MSCs)共培养时对各自分化的影响。方法:将2种细胞按接种数目分为5组,A组为NSCs∶MSCs=105∶0;B组为NSCs∶MSCs=105∶104;C组为NSCs∶MSCs=105∶105;D组为NSCs∶MSCs=104∶105;E组为NSCs∶MSCs=0∶105。培养7d后分别进行5-溴-2-脱氧尿嘧啶(BrdU)和神经元特异性烯醇化酶(NSE)、胶质细胞纤维酸性蛋白(GFAP)、微管相关蛋白2(MAP2)、半乳糖神经酰胺(GalC)免疫细胞化学双染色。结果:随着MSCs比例的提高,各组BrdU和NSE或MAP2的双阳性细胞表达差异有统计学意义(P<0.05),且B、C及D组分别与A组比较差异亦有统计学意义(均P<0.01);随NSCs比例的增高,B、C、D组NSE、MAP2和GFAP双阳性MSCs表达逐渐增高(均P<0.01)。结论:MSCs可以促进NSCs分化为神经元,而且在NSCs诱导下部分MSCs可以转化为神经元和星形细胞。

Objective：To study the differentiation of neural stem cell （NSCs） and bone marrow stromal cells （MSCs） when two kinds of cells were co-cultured. Methods：Two kinds of cells were divided into 5 groups by the inoculated number, （A） NSCs∶MSCs=105∶0; （B） NSCs∶MSCs=105∶104; （C） NSCs∶MSCs=105∶105; （D） NSCs∶MSCs =104∶105; （E） NSCs∶MSCs=0∶105. The cells were stained by 5-bromo-2-deoxyuridine （BrdU） and neuron-specific enolase （NSE）,glial fibrillary acidic protein（GFAP）,microtubule-associated protein（MAP2） and galactoceramide （GalC） double immunocytochemical staining after 7 days of culture. Results： The expressions of NSE and MAP2 were more according to the more MSCs （P〈0.01）. The proportion of double staining of NSE or MAP2 with BrdU was higher in group B,group C and group D than that of group A （P〈0.01）. The levels of NSE, MAP2 and GFAP were expressed only in MSCs of group B, C and D,and the expressions of NSE, MAP2 and GFAP increased with the increasing trend of NSCs （P〈0.01）. Conclusion： Bone marrow stromal cells can induce NSC differentiation into neurons and part of them can be induced the differentiation into neuron-like and astrocytes by NSCs.