诱导兔骨髓基质细胞向神经干细胞分化的初步实验研究

Preliminary Study on Induction and Differentiation of Rabbit's Bone Marrow Stromal Cells Into Neural Stem Cells

目的 :观察兔骨髓基质细胞体外培养的生长行为及增殖、分化情况 ,探讨由骨髓分离培养神经干细胞的可行性和有效性。方法 :无菌条件下行骨穿术 ,密度梯度离心分离获取骨髓基质细胞 ,利用多种增殖、分化诱导因子和神经干细胞培养液进行培养、分化诱导 ,用免疫细胞化学方法进行鉴定。结果 :骨髓基质细胞在体外培养中能形成NESTIN抗原阳性的细胞克隆团 ,进一步分化 ,部分细胞胞体增大并出芽 ,逐渐发育成为较成熟的长突起细胞 ,长突起相互连接、交织成网并建立纤维联系。结论 :骨髓基质细胞具有较强的自我更新能力和多向分化潜能 ,易于取材、体外培养和增殖 ,在一定诱导条件下可以生成神经干细胞。

Objective: To observe the growth, expansion and differentiation of the cultured rabbits' bone marrow stromal cells (BMSC) into neural stem cells and to assess the feasibility and validity of isolating and harvesting neural stem cells by this procedure. Methods: BMSCs were separated from other cells of bone marrow by density gradient centrifugation and cultured in neural stem cell medium supplemented with various neuro-nutrition factors, proliferation factors and differentiation-induced factors. Various bone marrow cells were defined immunocytochemically. Results: BMSC could be expanded and cloned when they were cultured in vitro. Part of the BMSCs showed Nestin positive. Further, some BMSCs formed small buds and developed gradually into relative mature cells with long projects that might connect with each other. Conclusion: This article shows that BMSC has the potential to renew and proliferate themselves and differentiate along certain direction, including differentiation into neural stem cells.