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Objective: isolated from human umbilical cord, cultured mesenchymal stem cells (mesenchymal stem cells, MSCs) to be identified, study of aplastic anemia (aplastic anemia AA) in patients with bone marrow and peripheral blood mononuclear cells (Mononucleae cell, MNC) The hematopoietic negative regulatory factor expression regulating role, to looking for easier to obtain and easy to cultivate and purification of MSC new sources and clinical treatment of AA, graft anti-host disease (GVHD) and improve the success rate of allogeneic hematopoietic stem cell transplantation (HSCT), etc. cytological methods and experimental evidence. Methods: A tissue culture method to train the MSC isolated from human umbilical cord (HUC); morphological and functional identification of the MSC; separated by density gradient centrifugation AA patients with bone marrow and peripheral blood mononuclear cells (MNC); HUC-MSC the AA patients MNC 1:10 a mixed culture, using real-time quantitative PCR (Real time PCR, RT-PCR) method were measured before and after the co-culture that 0h and 72h hematopoietic negative regulatory factor of TGF-β1, IFN-γ, TNF -α, T-bet mRNA expression changes. The measured data using SPSS17.0 software package for the analysis. Results: second generation HUC-MSC morphology typical long spindle change and growth characteristics of cultured comprehensive analysis of the results showed that the use of umbilical cord can be stable through MSC. Umbilical cord sources MSC and the AA patients bone marrow, peripheral blood MNC culture, co-culture before and after each factor concentration than the internal control β-actin and relative quantification of the mean and standard deviation were: T-bet: 0.255 ± 0.046 before co-cultured for 72h 0.201 ± 0.039, after the control group: 0.188 ± 0.023 (P gt; 0.05), the difference was not statistically significant; TGF-p1: co-culture 0.152 ± 0.041, co-cultured for 72h 0.321 ± 0.034, control group: 0.122 ± 0.024 ( P lt; 0.05), the difference was statistically significant, the concentration of a significant increase in the co-culture; TNF-α: co-cultured 0.634 ± 0.035, before co-cultured for 72h 0.198 ± 0.051, control group: 0.246 ± 0.043 (P lt; 0.05) the difference was statistically significant, concentration decreased after co-culture; IFN-γ: co-culture co-culture before 0.608 ± 0.066, 0.213 ± 0.037, control group: 0.178 ± 0.025 (P lt; 0.05), the difference was statistically significant, concentration decreased after co-culture. The significant difference between the experimental group and the control group and the experimental group (P lt; 0.05). Conclusion: Studies have shown can be successfully isolated from human umbilical cord tissue in the MSC, the MSC in morphological characteristics; HUC-MSC AA patients with bone marrow, peripheral blood MNC culture found HUC-MSC inhibit AA patients with T-cell expressed IFN-γ, TNF-α mRNA of levels at the same time raised the level of TGF-β1 mRNA, no significant effect on the mRNA levels of T-bet; before and after co-culture of iron deficiency anemia in patients with T-cell expression of TGF-β1, IFN-γ, no significant effect of TNF-α mRNA levels of T-bet. Umbilical cord derived MSC could by down-regulating IFN-γ and TNF-α levels at the same time raised the level of TGF-β1, and adjust the AA immune dysfunction, it may become a new method of treatment of AA MSC infusion.
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