骨髓基质细胞条件培养液中神经活性物质的提取与纯化

目的分离纯化骨髓基质细胞(bone marrow strolnal cells，BMSCs)条件培养液以获得某些神经活性物质。方法分离培养小鼠骨髓基质细胞并收集其培养液，超滤浓缩后采用Sephadex G-100层析，高效液相色谱分析(HPLC)和十二烷基硫酸钠-聚丙烯酰胺凝胶不连续电泳(SDS-PAGE)等技术分离其培养液中的蛋白组分并利用细胞培养技术验证其神经活性作用。结果骨髓基质细胞培养液经Sephadex G-100层析及HPLC分析，证实Ⅲ峰中的A峰和B峰对神经细胞生长有明显的促进作用，其相对分子质量分别为18000和14000。结论骨髓基质细胞条件培养液中相对分子质量为18000和14000两组分对神经元有营养活性。

Extraction and purification of neurotrophins from bone marrow stromal cell conditioned medium

Objective To isolate and purify neurotrophic active substances from bone marrow stromal cell (BMSCs) conditioned medium. Methods BMSCs were isolated from bone marrow of rats weighing 20 grams and cultured. The culture medium was then collected. Neurotrophic active substances in BMSCs conditioned medium were isolated and purified by means of Sephadex G-100 gel chromatography, HPLC, and SDS-PAGE. The neurotrophic bioactivity of isolated proteins was tested by cell culture. Results Sephadex G-100 gel chromatography and HPLC extracted two proteins of neurotrophic bioactivity from the BMSCs conditioned medium. One was at peak A with a molecular weight of 18KD, the other being at peak B with a molecular weight of 14KD. Culturing neurons from spinal cord anterior horn in the BMSCs conditioned medium containing these two proteins showed that growth of the neurons was promoted. Conclusion The proteins of 18 KD and 14 KD molecular weight in BMSCs conditioned medium have neurotrophic activity.