壳聚糖载体加载碱性成纤维细胞因子诱导成年大鼠骨髓间充质干细胞分化为神经细胞的研究

Differentiation of bone marrow mesenchymal stem cells of adult rat into nerve cells induced by chitosan carrier loaded with bFGF

摘要:

利用生物材料在体外将骨髓间充质干细胞诱导为神经元样细胞。方法利用全骨髓贴壁培养法提取骨髓间充质干细胞，并利用细胞表面抗原CD45 和CD90 进行鉴定。分为3 组，单纯壳聚糖组、加载了碱性成纤维细胞因子（bFGF）的壳聚糖组和单纯bFGF组。分别在3 和7 d 时利用免疫荧光细胞化学检测神经干细胞标记蛋白Nestin、幼稚神经元标记蛋白Tuj-1 等指标的表达情况。结果加载了bFGF 的壳聚糖处理细胞时，细胞在第3 天出现了大量的Nestin 阳性细胞，虽然在其他两组中也出现了Nestin 阳性细胞，但加载了bFGF的壳聚糖组比其他两组多，且差异有统计学意义（ p<0.05）。同样的，在诱导7 d时，Nestin 和Tuj-1 双阳性的细胞在加载了bFGF 组中的比例要比其他两组要多，且差异有统计学意义（ p<0.05）。结论加载有bFGF的壳聚糖能够将成年大鼠骨髓间充质干细胞高比例诱导分化为神经元样细胞。为骨髓间充质干细胞自体移植治疗神经退行性疾病和中枢神经系统提供了理论和实验依据。

Abstract:

To induce differentiation of bone marrow mesenchymal stem cells into neuron-like cells by biological material. Methods Using bone marrow adherent characteristic, bone marrow mesenchymal stem cells were extracted. Then the cells were identified by cell surface antigens CD45 and CD90. The cells were divided into three groups namely pure chitosan, chitosan-loaded bFGF group and bFGF group. The expressions of neural stem cell marker Nestin and neuron-like cell marker Tuj-1 were evaluated using immunocytochemical method on the 3rd and 7th d respectively. Results The proportion of Nestin-positive cells in the bFGF loaded group on the 3rd day was significantly higher than that in the other two groups ( p< 0.05). Similarly, the proportion of Nestin and Tuj-1 double positive cells in the bFGF loaded group was significantly higher than that in the other two groups on the 7th day of induction ( p< 0.05). Conclusions bFGF-loaded chitosan can induce differentiation of bone marrow mesenchymal stem cells of adult rat into neuron-like cells at a high proportion, which provides a theoretical and experimental evidence for autologous transplantation in the treatment of neurodegenerative diseases and central nervous system injury.