人AGM区基质细胞系的建立及其生物学特性  

Establishment of Stromal Cell Lines of Human Aorta-gonad-mesonephros (AGM) Region and Its Biological Characteristics

作　　者：陈惠芹[1] 张绪超[1] 黄绍良[1] 魏菁[1] 吴北燕[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1期20-23,28,共5页Journal of Sun Yat-Sen University:Medical Sciences

基　　金：国家自然科学基金(30300377);教育部博士点基金(20030558070);中国博士后科学基金资助项目(20003033432)

摘　　要：【目的】建立人主动脉鄄性腺鄄中肾(AGM)区基质细胞系,研究其生长特性及表面标志的表达。【方法】取孕30～35d的人胚胎分离培养AGM区基质细胞,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志和细胞外基质蛋白成分。【结果】人AGM区基质细胞呈成纤维样形态,指数生长期倍增时间约为16h。流式细胞仪检测结果显示CD31、CD34、CD45在人AGM区基质细胞表面为阴性表达,而CD29、CD44、CD105、CD166均为阳性表达。免疫荧光检测结果显示AGM区基质细胞表达细胞外基质蛋白Tenascin、Fibronectin、Laminin及α鄄SMA。【结论】人AGM区基质细胞系呈现造血组织特异的相关分子表达特征,可以作为研究造血发生机理的模式细胞。To establish stromal cell lines of human aorta-gonad-mesonephros (AGM) region and study the growth characteristics and hematopoiesis related molecules expression in these cells. Stromal cells of AGM region in human embryos of day 30-35 were isolated and cultured in vitro. Besides their growth characteristics were studied, hematopoiesis related molecules including surface and extracellular matrix proteins were analyzed respectively by flow cytometry and immunofluorescence methods. Stromal cells of human AGM region held the morphology of fibroblastoid cells. During logarithmic growth period, the cell population doubling time is about 16 h. By flow cytometry, surface markers CD31, CD34, CD45 were negatively expressed, while CD29, CD44, CD105, CD166 were positively detected on human AGM stromal cells. Through immunofluorescence assay, hematopoiesis related extracellular proteins such as tenascin, fibronectin, laminin and α-SMA were all positively expressed by stromal cells of AGM region. [Conclusion] These stromal cells of human AGM region have the characteristics of early blood-forming tissue specific molecules expression, and can be used as model cells for further hematopoiesis mechanisms research.

关 键 词：基质细胞 表达 表面标志 造血组织 细胞外基质蛋白 CD29 CD31 传代培养 性腺 免疫荧光检测 

分 类 号：R735[医药卫生—肿瘤] R331[医药卫生—临床医学]