长期体外扩增降低胎盘绒毛膜间充质干细胞的免疫调节功能  

Long Term In-vitro Expansion Reduces Immune Modulation Function of Placental Chorionic Villi Mesenchymal Stem Cells

作　　者：杨舟鑫 及月茹 韩之波[1,2] 王有为[1,2] 孟磊[1,2] 韩忠朝[1,2] 

机构地区：[1]中国医学科学院、北京协和医学院血液学研究所、血液病医院实验血液学国家重点实验室,天津300020 [2]中国医学科学院血液学研究所泰达生命科学技术研究中心,天津300457

出　　处：《中国实验血液学杂志》2013年第6期1552-1556,共5页Journal of Experimental Hematology

基　　金：国家重点基础研究发展计划"973计划"(编号2011CB964800);国家高技术研究发展计划"863计划"(编号2011AA020118)

摘　　要：本研究主要探讨长期体外培养的胎盘绒毛膜间充质干细胞的各项生物学活性和免疫调节功能的变化。显微镜检观察比较第3代和第9代胎盘绒毛膜的形态,用流式细胞术分析它们的免疫表型。把第3代和第9代CV-MSC与PHA活化的PBMNC共培养,ELISA方法检测其IFN-γ的分泌水平。实时定量PCR的方法检测CV-MSC细胞中COX-2,HGF和HLA-G的表达。结果显示:经过长期传代后,虽然CV-MSC的基本细胞形态和大部分表面标记如CD31,CD34,CD44,CD45,CD62L,CD73,CD90,CD105,CD117,CD151,CD235a,CD271和HLA-DR等都没有明显的变化,但其表面CD49d表达明显上调,长期传代后其免疫调节功能明显下降,免疫调节相关的分子COX-2和HGF的表达也略有下调,而HLA-G表达并未明显的变化。结论:长期体外扩增改变CV-MSC的CD49d的表达,并降低CV-MSC的免疫调节功能。The main aim of this study was to investigate the biological activities and immune modulation changes of chorionic villi mesenchymal stem cells（CV-MSC） after long term culture. The morphology of the CV-MSC of passage 3 and passage 9 were observed by micrscopy, and their phenotypes were detected by flow cytometry. CV-MSC of passage 3 and 9 were co-cultured with PHA-stimulated PBMNC, and IFN-γ concentration in culture medium was deteced by ELISA. The mRNA expression of COX-2, HGF and HLA-G in CV-MSC were detected by real-time PCR. The results showed that after long term culture, the CV-MSC kept the MSC morphology and most of the phenotypes including CD31, CD34, CD44, CD45, CD62L, CD73, CD90, CD105, CDl17, CD151, CD235a, CD271 and HLA-DR, while the CD49d was significantly up-regulated. Immune modulation ability of CV-MSC was reduced and the rnRNA expres- sion of COX-2 and HGF was down regulated after long term culture, but the expression of HLA-G did not found to be obvious change. It is concluded that the long term in vitro expansion changes the expression of CD49d and reduces im- mune modulation of CV-MSC.

关 键 词：长期培养 胎盘绒毛膜间充质干细胞 免疫调节 

分 类 号：R329.28[医药卫生—人体解剖和组织胚胎学]