胎肝造血期胎肝间充质干细胞的分离、培养及生物学特性  

Isolation, Culture, and Biological Characteristics of Mesenchymal Stem Cells from Fetal Liver in Fetal Liver Hematopoietic Stage

作　　者：吴北燕[1] 黄绍良[1] 陈惠芹[1] 张绪超[1] 魏菁[1] 

机构地区：[1]中山大学附属第二医院干细胞研究中心,广东广州510120

出　　处：《中山大学学报（医学科学版）》2005年第5期498-501,共4页Journal of Sun Yat-Sen University:Medical Sciences

基　　金：国家自然科学基金资助项目(30300377);教育部博士点基金资助项目(20030558070);中国博士后科学基金资助项目(2003033432);"十五"863计划资助项目(2003AA205008)

摘　　要：【目的】分离、培养人胎肝造血期胎肝间充质干细胞(MSC),研究其生长特性及表面标记的表达。【方法】取孕16～20周人胚胎肝脏,分离培养胎肝MSC,传代培养成系并检测其增殖能力,应用流式细胞术、免疫荧光方法检测其造血相关表面标记和细胞蛋白表达。【结果】人胎肝MSC呈成纤维样形态,指数生长期倍增时间约为16h,细胞增殖26.5倍。传代15次仍有94.18%的细胞处于G0/G1期。流式细胞仪检测结果显示人胎肝MSC表达CD29、CD44、CD105、CD106和CD166,不表达CD31、CD34、CD45,不表达与GVHD相关的HLA-DR、CD80、CD86、CD40、CD40L。免疫荧光检测结果显示胎肝MSC表达造血微环境细胞外基质蛋白Fibronectin、α-SMA及上皮细胞标记蛋白AFP和E-cadherin。【结论】人胎肝MSC具有胚胎造血组织和发育中的肝脏特异的相关分子表达,免疫原性弱,可以作为研究造血、胎肝发育机制的模式细胞。[Objective]To isolate and culture the mesenehymal stem cells （MSC） from fetal liver in fetal liver hematopoietie stage and study the growth characteristics and surface markers in these ceils. [Methods] MSC of fetal liver in human embryos of 16-20 weeks were isolated and cultured in vitro. Besides their growth characteristics were studied, expression of hematopoiesis related surface markers and proteins were analyzed by flow eytometry and immunofluorescenee methods. [ Results ] Fetal liver MSC held the morphology of fibroblastoid cells. During logarithmic growth period, the cell doubling time was about 16 h. The maximal cell number was increased by 26.5 times. 94.17% of fetal liver MSC in P15 remained in G0/G1 stage. By flow eytometry, surface markers CD31, CD34, CD45 were negatively expressed, either the GVHD related HLA-DR, CD80, CD86, CD40 and CD40L. However, CD29, CD44, CD105, CD106, and CD166 were positively detected. Through immunofluoresoenee assay, hematopoiesis related extracellular proteins fibronectin and α-SMA were positively expressed in fetal liver mesenchymal stem cells, as well as the epithelial marker protein AFP and Ecadherin. [Conclusions]Fetal liver MSC expressed some liver specific proteins as in fetal hematopoietic tissue, and in developing liver. It presents very weak immunogenicity, and can be used as model cells for further research in hematopoiesis and fetal liver ontogeny mechanisms.

关 键 词：胎儿 肝脏 间充质干细胞 造血发生 肝脏发生 

分 类 号：R329.1[医药卫生—人体解剖和组织胚胎学]