骨髓间充质干细胞促进中脑前体细胞增殖及定向分化的实验研究

目的 观察BMSCs对胚胎腹侧中脑前体细胞(VMP)体外扩增和定向分化的影响,并分析其可能的营养机制.方法 分别取胎龄11d大鼠胚胎VMP、成年大鼠BMSCs进行体外培养,并建立二者的共培养体系.体外扩增7 d的VMP分为对照组、BMSCs分化液组、BMSCs+VMP共培养分化液组,分别加入普通分化液、BMSCs分化液、BMSCs+VMP共培养分化液进行诱导分化.观察细胞生长情况.分化期末行免疫荧光染色,分析比较3组细胞中酪氨酸羟化酶(TH)阳性细胞占总细胞数的比例.结果诱导分化7 d后,对照组、BMSCs分化液组和BMSCs+VMP共培养分化液组中细胞数分别较培养前扩增(44.13±4.75)倍、(60.63±5.25)倍、(64.00±7.63)倍,TH阳性细胞比例分别为(18.76±5.20)%、(23.49±4.10)%、(28.08±5.42)%,比较差异有统计学意义(P<0.05).结论 BMSCs能够通过分泌营养因子有效促进VMP增殖并定向分化为多巴胺能神经元.

Bone marrow-derived mesenchymal stem cells inducing the proliferation and differentiation of ventral mesencephalic precursors

Objective To explore the potential neurotrophic effect of bone marrow-derived mesenchymal stem cells (BMSCs) on the proliferation and differentiation of ventral mesencephalic precursors (VMPs).Methods VMPs from E 11 inbred rat embryos and BMSCs from adult rats were cultured in 2 separate groups.Moreover,a co-cultured group was also established.Afterwards,the 3 different differentiation mediums obtained from these 3 defined groups were used to influence the differentiation procedures of normal VMPs that had amplification in vitro for 7 d.The growth of cells was observed;immunocytochemical staining was performed on these cells at the late differentiation phase.Relative yields of TH+ cells were calculated and compared.Results Seven d after the inducing differentiation,the total cell numbers multiplied about 44.13±4.75,60.63±5.25 and 64.00±7.63 folds in the VMP control group,the BMSCs group and the VMP+BMSCs co-cultured group,respectively,as compared with those before the differentiation.Correspondingly, the ratios of TH+ cells in the total cell population were (18.76±5.20)%, (23.49±4.10)% and (28.08±5.42)% in the VMP control group,the BMSCs group and the VMP+BMSCs co-cultured group,respectively; significant differences betwwen each 2 groups were found (P<0.05).Conclusion BMSCs can secret nutritional factors to promote the proliferation and committed differentiation of VMPs into dopaminergic neurons.