OECs无血清上清液体外诱导UB-MSCs向神经细胞分化

目的:观察大鼠嗅鞘细胞无血清上清液诱导脐血间充质干细胞向神经细胞分化的作用。方法:采用差速贴壁联合阿糖胞苷抑制法获得较高纯度的嗅鞘细胞,在最佳状态细胞无血清培养后取上清液。收集正常足月剖腹产胎儿的脐带血,经肝素抗凝,用淋巴细胞分离液分离脐血的单个核细胞,采用差速贴壁法纯化间充质干细胞,传第三代后用上述收集的嗅鞘细胞无血清上清液诱导,观察诱导分化过程,免疫组化鉴定分化结果。结果:用差速贴壁和阿糖胞苷抑制法可获得纯度高达95%的嗅鞘细胞,使用无血清嗅鞘细胞上清液培养脐血间充质干细胞后,发现能诱导脐血间充质干细胞向神经细胞形态分化,胞体呈椭圆形,伸出长突起。诱导7天后相差显微镜下可观察到神经干细胞、神经元和胶质细胞形态的细胞,与NSE、GFAP细胞免疫组化结果一致。结论:大鼠嗅鞘细胞无血清上清液有明显诱导脐血间充质干细胞分化为神经细胞的作用。

Differentiation of Human Umbilical Cord Blood Mesenchymal Stem Cells into Neural Cells Induced by Serum-free Conditioned Medium of OECs from Neonatal Rat

Objective To study the possibility and mechanism by which the human umbilical cord blood mesenchymal stem cells(MSCs) are induced to differentiate into neural cells in vitro by serum free condition medium of olfactory ensheasing cells(OECs) from neonatal rat.Methods Primary culture and purification of neonatal rat OECs in vitro with Nash differential adhesion and cytarabine inhibition method,extract serum-free condition medium of OECs when it is at the best situation.Human umbilical cord blood was collected from full term deliveries scheduled for cesarean section,all specimens were obtained sterilely with preservative-free heparin,the mesenchymal stem cells were isolated by lymphocyte separation medium,purified by Nash differential adhesion.After the passage 3 of the MSCs,add the serum free condition medium of OECs,then to observe the change of cell shape by microscop and immunocytochemistry.Results The OECs could be purified to about 95 percent purity by the method,MSCs displayed neuronal morphologies and there were long processes out from the cell body when the MSCs were induced by serum-free condition medium of OECs,and almost all human USB-MSCs appeared Neuron-like cells 2 weeks following induction.so as the number of immunocytochemistry cells labeled by neural markers NSE and GFAP.Conclusion The serum free condition culture medium of OECs can obvi ously induced human umbilical cord blood mesenchymal stem cells to differentiate into neural cells