论文部分内容阅读
目的:研究脑源性神经节苷脂诱导成年大鼠骨髓基质细胞定向分化为神经前体细胞的作用。方法:成年大鼠骨髓基质细胞传 代纯化后,分别在无血清、无外源性生长因子、含2%B27的 Neurobasal-A 培养基及含20%胎牛血清的 DMEM 培养基中添加脑源性 神经节苷脂作为诱导物,观察细胞的形态变化。结果:在无血清、无外源性生长因子、含2%1327的 Neurobasal-A 培养基中,脑源性神 经节苷脂能诱导骨髓基质细胞定向分化为神经前体细胞,其形态发生显著变化,胞体逐渐变小、折光性增强,突起逐渐增多、延长,部 分细胞间可见突起连接、细胞核和核仁;诱导第5 d 免疫细胞荧光染色显示,神经前体细胞标记物Nestin染色阳性,同时部分细胞可 见神经元特异性标记物 NSE 及星形胶质细胞标记物 GFAP 染色阳性,而少突胶质细胞标记物 Nogo-A 染色阴性。在含20%胎牛血 清的 DMEM(高糖)培养基中,脑源性神经节苷脂的这种诱导、分化作用明显被抑制,表现为细胞形态变化不明显,Nestin、NSE 及 GFAP 染色均为阴性。两组对照组细胞形态无变化、上述染色均为阴性。结论:脑源性神经节苷脂具有诱导骨髓基质细胞成为神经 前体细胞的作用,这种作用不需要外源性生长因子存在,血清可抑制这种作用,脑源性神经节苷脂在成体干细胞可塑性研究中有重要 价值。
AIM: To investigate the effect of brain derived gangliosides on the differentiation of adult rat bone marrow stromal cells into neural progenitor cells. METHODS: Adult rat bone marrow stromal cells were subcultured and purified. Brain derived cells were cultured in serum-free, non-exogenous growth factors, Neurobasal-A with 2% B27 and DMEM with 20% fetal bovine serum Gangliosides as inducers, observe the morphological changes of cells. RESULTS: Brain derived gangliosides induced differentiation of bone marrow stromal cells into neural progenitor cells in a serum-free, exogenous growth factor-free, Neurobasal-A medium containing 2% 1327, with significant morphological changes , The cell body gradually became smaller, the refractive index increased, and the number of protuberances increased gradually. Some of the cells showed prominent connections, nuclei and nucleolus. Fluorescent staining of immune cells on day 5 showed positive staining of Nestin in neural precursor cells, The neuron-specific marker NSE and astrocyte marker GFAP were positive for cells, whereas oligodendrocyte marker Nogo-A was negative. In DMEM (high glucose) medium containing 20% fetal bovine serum, the induction and differentiation of brain-derived ganglioside were significantly inhibited, showing no significant changes in cell morphology. The staining of both Nestin, NSE and GFAP Negative. There was no change in the cell morphology of the two control groups, the above staining were negative. CONCLUSION: Brain-derived gangliosides have the effect of inducing bone marrow stromal cells to become neural progenitor cells. This effect does not require the presence of exogenous growth factors. Serum inhibits this effect. Brain-derived gangliosides inhibit neuronal apoptosis in adult Stem cell plasticity research has important value.