摘要
建立体外稳定生长的人羊水干细胞(AFS)系并对其生物学特性作进一步鉴定。使用贴壁法分离培养羊水干细胞,通过扩增传代,建立体外稳定生长的人羊水干细胞系,采用RT-PCR检测干性基因的表达,流式细胞仪分析细胞表型,混合淋巴细胞培养检测对PBMC体外增殖的免疫调节作用并通过细胞计数研究其体外增殖潜能。成功建立三株体外稳定传代和生长的人羊水干细胞系,AFS表达干细胞特异性基因:Oct4、Sox2、Nanog、Klf-4、c-Mye和Rex-1;同时发现AFS高表达负性共刺激分子B7-H1、B7-H2、B7-H3、B7-H4、BTLA和不表达HLA-DR、CD40、CD80和CD86,AFS可抑制PBMC的体外增殖,不仅显示其具有低免疫原性,而且具有免疫调节作用。与此同时,AFS具有很高的体外增殖潜能。本研究建立的AFS系具有良好的干细胞生物学特性,可以作为一种来源充足的低免疫原性种子细胞用于组织工程的研究和应用。
This study aims to establish human amniotic stem cell（AFS）lines which can grow stably in vitro and may be used further for identification of their biological characteristics.By isolating and culturing AFS with adherent method,we established cell lines which could grow stably in vitro,and the stem cell gene markers were detected by RT-PCR,and we also analyzed the phenotype of AFS by flow cytometry.We have successfully established three AFS cell lines,and their influence on PBMC proliferation was examined by mixed lymphocyte culture and their proper proliferation in vitro by cell counting.The resuits showed that the AFS lines generally expressed stem cell-specific markers Oct4,Sox2,Nanog,Klf-4,c-Myc and Rex-1.Flow cytometry showed that the AFS lines highly expressed costimulatory molecules B7-H1,B7-H2,B7-H3,BT-H4 and BTLA but not HLA-DR,CD40,CD80 and CD86.AFS lines inhibited the proliferation of PBMC in vitro suggesting low immunogenicity and effect of immunoregulation.We also detected that the cells have a great potential in proliferation in vitro.Taken together,these AFS lines show good biological properties of stem cells and might be used as a source of abundant low immunogenic seed cells for research and application in tissue engineering.
出处
《现代免疫学》
CAS
CSCD
北大核心
2014年第5期397-402,共6页
Current Immunology
基金
国家自然科学基金(31000654)
973前期研究专题项目(2012CB722302)
关键词
羊水干细胞
干细胞
共刺激分子
amniotic fluid stem cell
stem cell
costimulatory molecule
