枸杞多糖诱导人脐血间充质干细胞向神经元样细胞分化的实验研究

目的:探讨枸杞多糖诱导人脐血间充质干细胞(MSCs)向神经元样细胞分化的可行性及其机制。方法:无菌条件下收集正常足月儿的脐带血,经肝素抗凝,用相对密度1.077的淋巴细胞分离液分离脐血的单个核细胞,用低糖DMEM培养基进行培养和纯化扩增。选取第3代细胞进行诱导实验,当传代细胞长满瓶底的80%以上时,先用含15?S和10ng/ml bFGF的DMEM完全培养基预诱导24小时,然后用不含血清含1g/L枸杞多糖的DMEM培养基诱导,光镜下观察细胞形态,用免疫组化技术检测细胞Nestin和NF的表达。结果:预诱导后MSCs没有变化,而经枸杞多糖诱导4h后细胞即出现形态学上的改变,细胞变成不规则形,立体感增强,从胞体伸出突起。免疫组化检测显示,细胞Nestin、NF呈阳性。结论:人脐血间充质干细胞经枸杞多糖诱导可转化为神经元样细胞,其诱导机制可能与枸杞多糖的抗氧化作用有关。

Study of Differentiation of Human Umbilical Cord Blood Mesenchymal Stem Cells Into Neuron-like Cells Induced by Lycium Barbarum Polysaccharine

Objective:To study the differentiation possibility and mechanism of human umbilical cord blood mesenchymal stem cells (MSCs) into neuron-like cells induced by lycium barbarum polysaccharine (LBP).Methods:Human umbilical cord blood was collected from mature neonates.All samples were obtained sterilely with 20 U/ml heparin.The cord mononudear cells were isolated with lymphocyte separation medium (density 1.077g/ml),then purified and expanded with slight sugar DMEM medium.The third passage of the expanded MSCs were pretreated with pre-induction medium consisting of DMEM with 15% FBS and 10ng/ml bFGF for 24 hours,then induced with induction medium consisting of DMEM without serum but 1g/L lycium barbarum polysaccharine.The morphological changes of the cells were observed under phase contrast microscope.The neuron specific markers,nestin and neurofilament (NF),were evaluated by indirect immunocyto-chemistry staining.Results:After induc- tion for 4 hours,some showed morphological changes obviously.Twenty-four hours later,the differentiated cells showed typical neuronal morphology.The expression of nesfin and NF were positive.Conclusion:Lycium barbarum polysaccharine may induce the human umbilical cord blood MSGs into neuron-like cells.The mechanism of such induction may be related with its anti-oxi- dation characteristics.