人脐带间充质干细胞、人脐带间充质干细胞源男性生殖细胞样细胞的免疫学特性  

Immune Properties of Human Umbilical Cord Wharton′s Jelly-Derived Mesenchymal Stem Cells and ItsDifferentiated into Male Germ-Like Cells

作　　者：林丽敏[1] 邱学荣[2] 邱晓燕[1] 唐秋灵[1] 王鸿武[1] 林晓波[1] 谢丽春[1] 罗梅娟[1] 马桂霞[1] 林广裕[1] 史雪川[1] 马廉[1] 

机构地区：[1]汕头大学医学院第二附属医院儿科,广东汕头515041 [2]惠州市中心人民医院儿科,广东惠州516001

出　　处：《实用儿科临床杂志》2011年第17期1343-1346,共4页Journal of Applied Clinical Pediatrics

基　　金：国家自然科学基金(31040057;81070478);广东省科技计划项目(2009B080701082);汕头市重点科技项目(汕府科[2010]480号)

摘　　要：目的研究人脐带间充质干细胞(huMSCs)及其诱导分化为男性生殖细胞样细胞后的免疫学特性,为进一步研究huMSCs的移植特性提供实验依据。方法分离培养huMSCs,在男性生殖细胞培养条件下定向诱导其分化为huMSCs源男性生殖细胞样细胞。采用FACScan流式细胞仪检测诱导前后huMSCs表面抗原CD40、CD40L、CD80、CD86。半定量反转录(RT)-PCR检测诱导前后huMSCs的人类白细胞抗原(HLA)-Ⅰ、HLA-DR基因表达水平。采用CCK-8法检测诱导前后的不同数量级huMSCs对混合淋巴细胞反应体系中经植物血凝素(PHA)激活的同种异体外周血淋巴细胞(PBMC)增殖的影响。采用酶联免疫吸附试验检测诱导前后huMSCs对经PHA活化的T细胞分泌干扰素-γ(IFN-γ)的影响。结果 HuMSCs在男性生殖细胞培养条件下能分化为男性生殖细胞样细胞,诱导前后的huMSCs均表达HLA-Ⅰ,不表达CD40、CD40L、CD80、CD86和HLA-DR。huMSCs能抑制经PHA激活的T淋巴细胞增殖,并抑制T淋巴细胞IFN-γ的分泌,但huMSCs源男性生殖细胞样细胞并不能抑制经PHA激活的T淋巴细胞增殖及IFN-γ的分泌。结论 HuMSCs在体外定向诱导为男性生殖细胞样细胞后,诱导后的细胞与huMSCs一样具有低免疫源性,同时huMSCs在体外具有免疫抑制性,但诱导获得的huMSCs源男性生殖细胞样细胞不能抑制T细胞的增殖。Objective To investigate the immune properties of human umbilical cord Wharton′s Jelly-derived mesenchymal stem cells（huMSCs） and huMSCs differentiated into male germ-like cells in vitro,in order to provide experimental data for future fundamental study on transplantation.Methods HuMSCs were isolated and cultured,and huMSCs were induced to differentiate into male germ-like cells in an artificial microenvironment in vitro.The expressions of CD40,CD40L,CD80,CD86 molecules on the surface of huMSCs and huMSCs differentiated into male germ-like cells were detected by flow cytometry.The expressions of human leucocyte antigen（HLA）-Ⅰ and HLA-DR were examined in both differentiated and undifferentiated huMSCs cells using reverse transcript-polymerase chain reaction（RT-PCR）.Inhibition of the proliferation of allogenic peripheral blood mononuclear cell（PBMC） stimulated by phytohemagglutinin（PHA）were detected with addition of different number of differentiated and undifferentiated huMSCs cells by mixed lymphocyte culture [cell counting Kit-8（CCK-8） met-hod],and detected the secretion of interferon（IFN）-γ,by PHA-activated T lymphocytes using enzyme linked immunosorbent assay（ELISA）.Results The morphology of huMSCs under artificial microenvironment induction gradually changed from fibroblast to slender spindles and formed a ＂tadpole-like＂shape.Both differentiated and undifferentiated huMSCs cells expressed HLA-Ⅰ but didn′t express CD40,CD40L,CD80,CD86,HLA-DR.Mixed lymphocyte assays showed only undifferentiated huMSCs cells could suppress the proliferation of allogenic PBMCs.HuMSCs could inhibit the growth of T lymphocytes cell by PHA-activated and the secretion of IFN-γ were detected,while the male germ-like cells could′t.Conclusions HuMSCs can be differentiated into male germ-like cells in vitro,the male germ-like cells and huMSCs possess weak immunogenicity,huMSCs also perform immunoregulatory functions by suppressing T lymphocyte proliferation in vitro,but the male germ-like c

关 键 词：人脐带 间充质干细胞 诱导 免疫原性 免疫调节 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]