人脐带间充质干细胞对再生障碍性贫血患者Treg细胞的影响  

Effect of human umbilical cord mesenchymal stem cells on Treg cells with aplastic anemia patients

作　　者：柴晔[1] 刘瑛[1] 孙媛媛[2] 张连生[1] 曾鹏云[1] 吴重阳[1] 岳玲玲[1] 李莉娟[1] 熊彬[1] 

机构地区：[1]兰州大学第二医院血液科,兰州730000 [2]甘肃省人民医院儿科,兰州730000

出　　处：《中国免疫学杂志》2011年第9期837-839,共3页Chinese Journal of Immunology

基　　金：兰州大学医学科研基金项目(LZUYX200734)

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。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.

关 键 词：再生障碍性贫血 脐带间充质干细胞 调节性T细胞 

分 类 号：R392.12[医药卫生—免疫学]