【摘要】 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。更多还原

【Abstract】 Objective:To study the effect of human umbilical corb mesenchymal stem cells(hUC-MSC)on Treg cells with aplastic anemia(AA) patients,and explore their possibly therapeutic action for AA.Methods:The seventh passage of hUC-MSC of serial subcultivation were got,and the peripheral blood mononuclear cells of AA(AA-PBMC) were isolated by density gradient centrifugation.The experiment was divided into three groups:blank group(hUC-MSC),control group(AA-PBMC) and experimental group(hUC-MSC were cocultured with AA-PBMC at 1∶10,1∶1 and 10∶1 for 7days respectively).The relative expression of Foxp3mRNA(Foxp3/β-actin) was measured by RT-PCR.The proportion of Treg cells(CD4+CD25+Foxp3+)in peripheral CD4+T cells was analyzed by 3-color flow cytometry.Results:hUC-MSC didn’t express Foxp3mRNA and there were not Treg cells.The relative expression of Foxp3 mRNA and the proportion of Treg cells in peripheral CD4+T cells of experimental groups were higher than that of control group,and the difference was more significant when hUC-MSCs and AA-PBMC mixed at the ratio of 10∶1.Conclusion:hUC-MSC could increase the proportion of Treg cells in peripheral CD4+ T cells of aplasic anemia patients in vitro,which supplies theoretical foundation for clinical therapy.As their widespread sources and having been industrialized at present,hUC-MSC have broad perspective of clinical application.更多还原