【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。更多还原

【Abstract】 【Objective】To isolate and culture the mesenchymal stem cells (MSC) from fetal liver in fetal liver hematopoietic stage and study the growth characteristics and surface markers in these cells.【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 cytometry and immunofluorescence methods【.Results】Fetal liver MSC held the morphology of fibroblastoid cells. During logarithmicgrowth 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 cytometry, surface markers CD31, CD34, CD45were negatively expressed, either the GVHD related HLA-DR, CD80, CD86, CD40 and CD40L. However, CD29,CD44, CD105, CD106, and CD166 were positively detected. Through immunofluorescence assay, hematopoiesisrelated 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 liverspecific 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.更多还原