Insulin-like Growth Factor -1 Induces Differentiation Of Human Umbilical Cord Blood-derived Mesenchymal Stem Cell To Oligodendrocytes-like Cells

【Objective】Isolating,proliferating the mesenehymal stem cells(MSCs) from human umbili- cal cord blood in vitro,and inducing MSCs differentiation to oligodendrocytes by using insulin-like growth factor -1, neonatal rats O2A progenitors supernatants.To getting more oligodendrocytes provide the bases of oligodendrocyte related disease's research.【Methods】1. Human umbilical cord blood was collected in asepsis condition,isolated by density gradient centrifugation,the isolated mononuclear cells were cultured in vitro, DMEM culture medium containing 20% of fetal bovine seurm was used as the culutre system. For cell surface antigen phenotpying,third Passage cells were detached with fluorescein antibodies and analyzed with flow cytometer and were tested for their diffrentiation potentiality to lipoblast.2. To induce differentiation of MSCs by using basic fibroblast growth factor, insulin-like growth factor -1, neonatal rats O2A progenitors supernatants. The cell differentiation rate after the induction were detacted by the laser confocal microscope and Western blot method.【Results】1. The third passage cells were all spindle-like shape,grow directional showing parallel venation and reached 70-80% confluence after 3-4 weeks.Flow cytometric analysis demonstrated the cells were positive forCD44 and CD105 but negative for CD34 and CD45.The MSCs can be successful differentiated into adipogenic lineage.2.The inducing ability of 10ng/mL IGF-1 have significant difference to the control (P <0.05),the others have no significant difference (P> 0.05).3. There was no significant difference between Insulin-like growth factor -1, neonatal rats O2A progenitors supernatants of the amount of Galc (P> 0.05); the amount of Galc of the neonatal rats O2A progenitors supernatants and insulin-like growth factor-1 group have significant difference to using insulin-like growth factor-1 or neonatal rats O2A progenitors supernatants alone (P< 0.05).【Conclusion】1 Isolating by density gradient centrifugation and cultured in vitro, can collect pure human umbilical cord blood .2 Human umbilical cord blood can be differentiated to oligodendroglia in the condition of bFGF,IGF-1,but the differentiation rate is low and its biologycal activity is still unknow.3 The differentiation rate can be influence by the inductor.The experiment showed that 10ng/mL bFGF,10ng/mL IGF-1 is the appropriate concentration,and neonatal rats O2A progenitors supernatants can improve the induction rate.