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中文摘要：
目的观察与大脑皮质神经元共培养的骨髓基质细胞（BMSCs）经诱导分化成神经元样细胞后，与脑皮质神经元之间形成功能性突触的情况。方法无菌条件下取绿色荧光蛋白（GFP）转基因小鼠骨髓，用贴壁筛选法体外培养获得GFP转基因小鼠BMSCs（GFP-GM-BMSCs），在体外培养、扩增、纯化。取第3代GFP-GM-BMSCs，种植到源于小鼠大脑的原代皮质神经元和胶质细胞中，培养介质为加有20ng／mL表皮生长因子（EGF）、20ng／mL碱性成纤维细胞生长因子（bFGF）的无血清培养基（Neurobasal-A＋2％B27），体外模拟建立细胞移植的共培养体系。共培养第10天，利用FM1-43荧光染料染色活动突触小泡的特性，通过荧光显微镜观察共培养的两种细胞之间形成的突触。结果与神经元共培养的GFP-GM-BMSCs在含有EGF、bFGF的无血清培养基中7d后分化为神经元样细胞。共培养10d后，FM1-43染色阳性的突触囊泡明显增加，主要位于神经元样细胞胞体、突起及其末端结构上。结论在体外模拟细胞移植共培养体系中，分化自GFP-GM-BMSCs的神经元样细胞能与神经元之间形成突触样连接。
英文摘要：
Objective To investigate the establishment of synapses between the cortical neurons and the neuron-like cells differentiated from the marrow stromal cells （BMSCs） in a simulated transplantation system in vitro. Methods The BMSCs from green fluorescent protein （GFP） transgenic mice （GFP-GM-BMSCs） were isolated, cultured and purified in vitro. The third passage of GFP-GM-BMSCs were co-cultured with primary cultured cortical neurons and glial cells in a simulated transplantation system in serum-free medium containing 2% B27 supplemented with 20 ng/mL basic fibroblast growth factor （bFGF） and 20 ng/mL epidermal growth factor （EGF）. On day 10 of the co-culture, FM1-43, a fluorescent dye specific to active synaptic vesicles, was used to observe synapses formation between the cells under fluorescence microscope. Results The GFP-GM-BMSCs co-cultured with the neural cells in the serum-free medium containing bFGF and EGF differentiated into neuron-like cells 7 days after the co-culture. On day 10 of the co-culture, FM1-43 dye-positive synaptic vesicles were found in the cell culture, locating mostly in the cell body, processes and terminal structures of the neuron-like cells. Conclusions The neuron-like cells derived from GFP-GM-BMSCs can form synapses with the cortical neurons in the simulated cell transplantation system in vitro.
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