体外诱导骨髓基质细胞向神经干细胞和成熟神经细胞分化的实验研究

IN VITRO-INDUCED DIFFERENTIATION OF BONE MARROW STROMAL CELLS INTO NEURAL STEM CELLS AND MATURE NEURAL CELLS

探索骨髓基质细胞(BMSCs)分化为神经干细胞和成熟神经细胞的可行性 ,为BMSCs在神经科学领域的应用提供依据。以犬的BMSCs为实验对象 ,利用bFGF、RA、GDNF等作为增殖或分化诱导因子 ,对各阶段细胞进行免疫细胞化学鉴定。结果发现 ,增殖培养2 4～ 72h见细胞分裂像和克隆单位 ;诱导培养 3天 ,部分细胞开始表达NSE或GFAP ,继续增殖培养仍可见细胞克隆 (干细胞特性 ) ;诱导培养第 10天有成熟神经细胞形成 (成分鉴定证实 )。说明BMSCs在体外培养条件下 ,经过多种因子的“程序性”作用 ,可以向神经干细胞、成熟神经元及胶质细胞方向分化。提示BMSCs可作为“种子细胞”在神经科学领域加以利用。

To investigate the feasibility of in vitro inducing differentiation of bone marrow stromal cells (BMSCs) into neural stem cells and mature neural cells, and to offer reference for the application of BMSCs in the field of neuroscience, BMSCs were acquired from the marrow of dogs. Basic fibroblast growth factor (bFGF), all trans retinoic acid (RA) and glial cell line derived neurotrophic factor (GDNF) etc were used as proliferation or differentiation factors. Immunocytochemistry was employed to identify the cells at every culture stage. When BMSCs were proliferatively cultured for 24～72h, cleavage phase and cluster like clones appeared. On the 3rd day, some of the BMSCs derived cells started to express neuron specific enolase (NSE) or glial fibrillary acidic protein (GFAP). The same stage cells could be cloned, that is one of the characters of stem cells, when they were cultured in the proliferation medium. The neural cell modality like cells appeared on the 10th day after adding inducing factors into culture medium, which were proved by testing the express of NSE and GFAP. BMSCs can differentiate into neural stem cells and mature neural cells in vitro, and can be used as 'seed cells' in the field of neuroscience.