人脐带间充质干细胞对异体人外周血单个核细胞的免疫调节作用  

Immunomodulatory effect of human umbilical cord mesenchymal stem cells on allogeneic human peripheral blood mononuclear cells

作　　者：陈耐寒 赵仁彬[2] 孙鷖 李丽 郑永钦 左荣霞 张进萍 赵倩 沈涛 撒亚莲 CHEN Nai-han;ZHAO Ren-bin;SUN Yi;LI Li;ZHENG Yong-qin;ZUO Rong-xia;ZHANG Jin-ping;ZHAO Qian;SHEN Tao;SA Ya-lian(Affiliated Hospital of Kunming University of Science and Technology,Center for Clinical Medicine Research,The First People′s Hospital of Yunnan Province,Yunnan Provincial Key Laboratory for Birth Defects and Genetic Diseases,Yunnan Provincial Key Laboratory of Clinical Virology,Kunming 650032,Yunnan Province,China)

机构地区：[1]昆明理工大学附属医院云南省第一人民医院临床医学研究中心云南省出生缺陷与遗传病研究重点实验室云南省临床病毒学重点实验室,云南昆明650032 [2]云南省第一人民医院国家临床重点专科血液科,云南昆明650032 [3]云南省第一人民医院检验科,云南昆明650032

出　　处：《中国生物制品学杂志》2022年第7期802-807,共6页Chinese Journal of Biologicals

基　　金：云南省卫生厅内设机构资助项目(2017NS228,2018NS261);云南省卫生科技计划项目(L-2019003)。

摘　　要：目的探讨人脐带间充质干细胞(human umbilical cord mesenchymal stem cell,hUC-MSCs)对异体人外周血单个核细胞(peripheral blood mononuclear cells,PBMCs)增殖及其调节性T细胞(regulatory T cells,Treg)亚群的影响。方法用组织块贴壁法培养原代hUC-MSCs,经消化酶传代培养扩增细胞,获得第4代hUC-MSCs;经Ficoll密度梯度离心法获取PBMCs。将hUC-MSCs与PBMCs以1∶5和1∶10的比例共培养,设单独培养的PBMCs为对照组,在培养体系中添加25 ng/mL植物血凝素(phytohaemagglutinin,PHA),培养5 d;采用全自动血液分析仪检测PHA活化PBMCs数量,流式细胞仪检测CD4^(+)CD25^(+)CD127d^(dim/-)Treg细胞表达水平。结果hUC-MSCs为贴壁生长的梭形细胞,具有分化为成脂肪样细胞、成骨样细胞和成软骨样细胞的潜能。hUC-MSCs与PBMCs以1∶5和1∶10比例共培养5 d,悬浮细胞数均少于单独培养组,hUC-MSCs抑制PBMCs增殖率分别为(81.54±5.10)%和(43.37±3.35)%;hUC-MSCs促进CD4^(+)CD25^(+)CD127d^(dim/-)Treg细胞亚群增殖,上调效率分别为(41.73±4.66)%和(19.39±5.62)%,高于单独培养组。结论hUC-MSCs在体外对PHA活化的异体人PBMCs有免疫负调节作用。Objective To investigate the effects of human umbilical cord mesenchymal stem cells(hUC-MSCs)on proliferation and regulatory T cell(Treg)subsets of allogeneic human peripheral blood mononuclear cells(PBMCs).Methods Primary hUC-MSCs were cultured by tissue block adherent method then subcultured and proliferated by enzyme digestion,of which the passage 4 was subjected to Fiucoll density gradient centrifugation to obtain PBMCs.The hUC-MSCs and PBMCs were co-cultured at ratios of 1∶5 and 1∶10 respectively,using PBMCs cultured alone as control.The culture system was added with 25 ng/mL phytohaemagglutinin(PHA)and cultured for 5 d.The count of PHAactivated PBMCs was determined by fully automatic blood analyzer,while the expression level of CD4^(+)CD25^(+)CD127^(dim/-)Treg cell subsets by flow cytometry.Results The hUC-MSCs were spindle cells in adherent growth,with potentials of differentiation to adipocyte-like,osteoblast-like and chondroblast-like cells.The counts of hUC-MSCs co-cultured with PBMCs at ratios of 1∶5 and 1∶10 for 5 d were less than those cultured alone,while the proliferation-inhibiting rates of PBMCs were(81.54±5.10)%and(43.37±3.35)%respectively.However,hUC-MSCs co-cultured with PBMCs at ratios of 1∶5 and 1∶10 promoted the proliferation of CD4^(+)CD25^(+)CD127^(dim/-)Treg cells,which increased the percentages of the latter by(41.73±4.66)%and(19.39±5.62)%respectively as compared with those in control group.Conclusion The hUC-MSCs showed negatively immunomodulatory effect in vitro on the allogeneic human PBMCs activated by PHA.

关 键 词：人脐带间充质干细胞 外周血单个核细胞 调节性T细胞 免疫调节 

分 类 号：R392.12[医药卫生—免疫学]