BMP-4在人AGM区基质细胞的表达  

Expression of BMP-4 in Stromal Cells In Vitro Derived from Human Aorta-Gonad-Mesonephros Region

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

机构地区：[1]中山大学附属第二医院干细胞研究中心

出　　处：《中国实验血液学杂志》2007年第4期772-775,共4页Journal of Experimental Hematology

基　　金：国家自然科学基金资助项目(编号30300377);教育部博士点基金资助项目(编号20030558070)

摘　　要：本研究旨在观察BMP-4在人胚胎主动脉-性腺-中肾(AGM)区基质细胞的表达,为建立含AGM区基质细胞的诱导胚胎干细胞造血分化培养体系提供理论和实验依据。体外传代培养人胚胎AGM区基质细胞(hAGM S1-S5)及取自同期胚胎的躯干成纤维细胞(hFT),采用RT-PCR方法在mRNA水平分析BMP-4在hAGM S1-S5的表达情况,并应用ELISA方法测定BMP-4在hAGM S1-S5细胞培养上清液中的分泌水平,同时以hFT细胞作为对照。结果表明:体外培养中的hAGM S1-S5细胞形态学表现主要为成纤维间质细胞样,少量呈内皮样,是早期胚胎AGM区来源的异质细胞群。hAGM S1-S5细胞均表达BMP-4 mRNA,hFT不表达BMP-4 mRNA。hAGM S1-S5细胞培养上清液中可测得分泌的BMP-4,其分泌水平(pg/ml)分别为55.98±11.20、61.16±9.39、76.26±11.31、86.38±18.34、85.39±21.22,与hFT对照组(16.76±7.04pg/ml)相比有显著性差异(p<0.05),hAGM S1-S5组间比较BMP-4水平无显著性差异(p>0.05)。结论:体外培养中的人AGM区基质细胞表达BMP-4,与AGM区基质细胞支持造血干细胞原始发生的功能相关。The objective of this study was to investigate the expression of BMP-4 in stromal cells in vitro derived from human aorta-gonad-mesonephros （AGM） region, Stromal cells derived from human AGM region （hAGM S1 - S5 ） and fibroblasts derived from human fetal trunk （ hFF ） were cultured in vitro, RT-PCR was used to analyze the expression of BMP-4 in hAGM S1 - S5 stromal cells at mRNA level, And BMP-4 level was detected in the supernatant of hAGM S1 - S5 stromal cells by ELISA assay, hFF cells were used as control group, The results showed that the heterogenous hAGM S1 -S5 stromal cells displyed shapes of fibroblast-like and endothelial-like cells, hAGM S1 - S5 stromal cells expressed BMP-4 mRNA, but fetal trunk fibroblasts （hFF） did not express BMP-4 mRNA, In the supernatant of hAGM S1 - S5 cells, BMP-4 could be detected by ELISA assay ana its levels were statistically higher than that in hFF group （p 〈0, 05 ）, while there was no significant difference between groups of hAGM S1 - S5 （p 〉0.05 ）, It is concluded that human AGM-derived stromal cells in vitro express BMP-4, and the establishment of a new culture system based on the feeder cells of AGM stroma would promote the differention of embryonic stem cells into hematopoietic stem cells at a high proportion.

关 键 词：BMP-4 主动脉-性腺-中肾区 基质细胞 

分 类 号：R322.2[医药卫生—人体解剖和组织胚胎学]