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目的:研究人脐带间充质干细胞(hUC-MSC)对再生障碍性贫血患者Treg细胞的影响,探讨其对再生障碍性贫血(AA)可能的治疗作用。方法:取传代培养第7代的hUC-MSC,密度梯度离心法分离AA-PBMC,分为3组:空白组(hUC-MSC),对照组(AA-PBMC),实验组(分别以1∶10、1∶1及10∶1的比例将hUC-MSC与AA-PBMC共培养7天),RT-PCR法测定Foxp3mRNA的相对表达量(Foxp3/β-actin),3色流式细胞仪测定Treg细胞(CD4+CD25+Foxp3+)占外周血CD4+T细胞的比例。结果:hUC-MSC不表达Foxp3mRNA,无Treg细胞存在。实验组Foxp3mRNA相对表达量、Treg细胞占外周血CD4+T细胞比例高于对照组(P<0.05),hUC-MSC与AA-PBMC比例为10∶1时,差异更显著(P<0.01)。结论:人脐带间充质干细胞体外可增加再生障碍性贫血患者Treg细胞占CD4+T细胞比例,为临床治疗提供新的理论依据;其来源广泛,目前已产业化,具有广阔的临床应用前景。
Objective: To investigate the effect of human umbilical cord mesenchymal stem cells (hUC-MSCs) on Treg cells in patients with aplastic anemia and to explore its possible therapeutic effect on aplastic anemia (AA). Methods: The hUC-MSCs of passage 7 were passaged and the AA-PBMCs were separated by density gradient centrifugation. The cells were divided into 3 groups: hUC-MSC group, AA-PBMC group, HUC-MSCs were co-cultured with AA-PBMC for 7 days in a ratio of 10: 1 and 10: 1. The relative expression levels of Foxp3 mRNA and Foxp3 mRNA were determined by RT-PCR and flow cytometry Treg cells (CD4 + CD25 + Foxp3 +) in peripheral blood CD4 + T cell ratio. Results: hUC-MSC did not express Foxp3 mRNA and no Treg cells existed. The relative expression level of Foxp3 mRNA in experimental group was higher than that in control group (P <0.05). The ratio of Treg cells to peripheral blood CD4 + T cells was significantly higher than that of control group (P <0.01). Conclusion: Human umbilical cord mesenchymal stem cells can increase the proportion of Treg cells in CD4 + T cells in patients with aplastic anemia in vitro, which provides a new theoretical basis for clinical treatment. The umbilical cord mesenchymal stem cells have a wide range of sources and are currently industrialized and have broad clinical application prospects.