Human Umbilical Cord Mesenchymal Stem Cells In Patients With Aplastic Anemia Hematopoietic Regulation Of Negative Regulatory Factors

Objective:Isolated from human umbilical cord, cultured mesenchymal stem cells (MSCs) and to be identified to study the effect of aplastic anemia (AA) bone marrow or peripheral blood mononuclear cells part of hematopoietic negative regulatory regulation of the expression.Inorder to find more accessible and easy to culture and purification of a new souce of MSC and clinical treatment of aplastic anemia, graft versus host disease(GVHD) and enhance allogeneic hematopoietic stem cell transplantation(HSCT) success rate of cytological methods and provide experimental evidence.Methods:Using tissue culture from human umbilical cord (HUC) were isolated and cultured MSCs; by morphology and function of MSC comprehensive identification; by density gradient centrifugation of bone marrow in patients with AA, peripheral blood mononuclear cells; the HUC-MSC mononuclear cells in patients with AA ratio of 1:10 mixed culture, with real-time quantitative PCR (RT-PCR) were measured by semi quantitative before and after co-culture (0h and72h), cytokine TGF-β1, IFN-γ, TNF-α, T-bet in the mRNA expression level. The measured data were analyzed using SPSS 17.0 software.Results:The cultured for 2 generations of HUC-MSC with spindle morphology and growth characteristics of a typical change, integrated analysis showed characters using umbilical cord can get a stable MSC.MSC from umbilical cord and bone marrow of patients with aplastic anemia,peripheral blood mononuclear cells were cultured,the concentration of each factor before and after co-culture than the internal referenceβ-actin were auantified after the number and relative standard deviation are:T-bet:The former co-culture was 0.255±0.046,after 72h co-culture was 0.201±0.039, control group was:0.188±0.023 (P> 0.05), the difference was not statistically significant;TGF-β1:The former co-culture was 0.152±0.041, after 72h co-culture was 0.321±0.034, control group was:0.122±0.024 (P<0.05), the difference was statistically significant after co-culture was significantly increased;TNF-a:The former co-culture was 0.634±0.035, after 72h co-culture was 0.198±0.051, control group was:0.246±0.043 (P<0.05), the difference was statistically significant, decrease in concentration after co-culture;IFN-γ:The former co-culture was 0.608±0.066, after 72h co-cultured was 0.213±0.037, control group was:0.178±0.025 (P<0.05), the difference was statistically significant, concentration was decreased after co-culture.Experimental group and control group, the experimental group were significantly different (P<0.05).Conclusion:The studies have shown that human umbilical cord tissue from the umbilical cord successfully isolated MSC, it has the morphological characteristics similar to bone marrow MSC; the HUC-MSC and AA bone marrow, peripheral blood mononuclear cells were cultured and found that inhibition of AA patients HUC-MSC T cells expressed IFN-y, TNF-a in the mRNA level also increased the mRNA levels of TGF-β1 on the mRNA levels of T-bet had no significant effect; co-culture in patients with iron deficiency anemia before and after the T cells express TGF-β1, IFN-γ, TNF-α, T-bet in the mRNA levels were not significantly affected. MSC from the umbilical cord may be passed down IFN-y, TNF-a levels also increase the level of TGF-β1, and conditioning AA immune disorder, so the infusion of MSCs may become a new method of treatment of AA.