维生素C对骨髓基质细胞的诱导分化作用

Effect of Ascorbic Acid on Bone Marrow Stromal Cells differentiated into Neurons

目的观察维生素C对骨髓基质细胞(Bone marrow stromal cells,BMSCs)向神经元方向分化的作用及对其递质分泌的影响。方法采用percoll液分离大鼠骨髓基质细胞,体外扩增。当第二或第三代细胞生长达到60％～70％融合时,加入不同浓度的维生素C,分别于不同时间观察其形态学改变,取诱导后的细胞作免疫细胞化学鉴定,取诱导后的诱导液作HPLC测定其中的单胺类递质的含量。结果用不同浓度的维生素C可使骨髓基质细胞表现出典型的神经元样细胞的形态。免疫组织细胞化学鉴定显示,诱导后的细胞表现为NSE阳性,GFAP阳性。HPLC测定结果显示,用维生素C诱导后,诱导液中的肾上腺素和去甲肾上腺素的含量明显增多。结论维生素C可诱导骨髓基质细胞向神经元样细胞方向分化,并且可增加其中单胺类递质的释放。

Objective To observe ascorbic acid effect on bone marrow stromal cells differentiated into neurons and transmitters secreted.Methods BMSCs were separated from rat bone marrow.When the cells（from Passage2 or Passage 3）confluent about 60% -70%,the media were replaced with ascorbic acid （vitamin C,Vc）.Observed the morphology of the subpopulation cells of BMSCs at different time.Cells were fixed for immunocytochemistry. Detected the transmitters of catecholamine（CA） in medias by HPLC.Result After seven hours induced by 50 μ g/ mL Vc,the cells just transformed.After 3 hours induced with 100-200 μg/mL Vc,small cells displayed typical neuronal morphologies,but their apoptotic speed was quick.Flattened cells＇ transformation was slow.Treated by 500 μg/mL Vc,the cells transformation was faster than the other doses.The differentiated cells expressed GFAP and MAEThe content of N,NE increased in the induced medias.Conclusion Ascorbic acid could induce BMSCs to differentiating into neuron and could increase secreting of E and NE.