The Research For The Differentiation Potential Of Umbilical Cord Blood-derived Mesenchymal Stem Cells

Objective: Mesenchymal stem cells(MSCs) are adult stem cells and have differentiationpotentials，which are capable of differentiating into cells of different mesenchymal tissue lineagessuch as bone, cartilage, and adipose tissues. In recent study, it has been found that mesenchymalstem cells exsit in umbilical cord blood. As a new resource of stem cells，these mesenchymal stemcells are capable to repair the tissue defect in stem cells research，such as myocardiac infarction,due to their advantages such as extensive resources，easy to acquire, multi-differentiation potential.Moreover, they can be used to prophylaxis of graft versus host diseases (GVHD) due to lowerimmunogenicity. However,the quantity of them is very low in umbilical cord blood and it isextremely difficult to culture them in vtrio. Du to a great many controversies among the researchesof differentiations in UCB-MSCs comparing to that of BM-MSCs，We need to further study thedifferentiation potiential of UCB-MSCs. This study was to improve the successful rate of culturewith optimization of Culture in vitro and study further the potentials of adipocyte differentiationMethod:1. The volume of umbilical cord blood exceeded80ml and the UCB-MSCs samples wereobtained with histopaque density separation. Optimazition of the medium by adding tosuitable fetal bovine serum for improving the rate of culture.2. When the cells were subcultured to the4th or5th passage, they were evaluated by the flowcytometric staining with anti-body. RT-PCR was used to test the gene expression in CXCR4，SDF-1，CD54，CD49d of UCB-MSCs.3. UCB-MSCs were incubated in adiocyte differentiation medium.Results: The successful rate of primary cuture is high in the medium which contianed15%fetalbovine serum. These cells were negative for HLA-ABC，HLA-DR，CD13，CD14，CD31， CD106and postive for CD29，CD44，CD59，CD90，CD73，CD105，CD166. It is positivefor expression of gene in CXCR4,SDF-1,CD54,CD49d. After incubation in adiocytesdifferentiation medium, they differentiated into adipocyte cells.Conclusion: DMEM-low glucose medium containing15%fetal bovine serum is suitablemedium for the primary cutlure of UCB-MSCs in comparision with other mediums.We provedthat the cells we obtained were the UCB-MSCs according to the morphology of cells and thephenotype of cells suface and gene expression of adhesion molecular. The UCB-MSCs candifferentiate into adipocyte under the adipocyte differentiation medium.