人类胚胎干细胞向造血干细胞分化的微环境研究  

Microenvironment for the hematopoietic differentiation of human embryonic stem cells

作　　者：王建[1] 林戈[1] 赵惠萍[1] 卢光绣[1] 

机构地区：[1]中南大学生殖与干细胞工程研究所,人类干细胞国家工程研究中心,湖南长沙410078

出　　处：《南方医科大学学报》2009年第6期1119-1122,共4页Journal of Southern Medical University

基　　金：国家教育部博士点基金(20030533002);国家自然科学基金(30300119)

摘　　要：目的探讨胎肝基质细胞和骨髓基质细胞在诱导胚胎干细胞向血液血管干细胞分化中的作用,分析基因表达差异。方法通过将分化4d的EBs分别接种到以下两组中:(1)胎肝基质细胞层+细胞因子;(2)骨髓基质细胞层+细胞因子;诱导结束后,流式细胞术检测各组中血液血管干细胞标志KDR阳性细胞数以及造血干细胞标志CD34阳性细胞数,计数细胞分化过程中形成的造血干细胞样的集落数,比较诱导分化效果;收集胎肝基质细胞和胚胎骨髓基质细胞,采用cDNA芯片分析基因表达差异。结果流式细胞术检测发现,两组中KDR+细胞的比例为分别为:(1.06±0.20)%、(8.8±1.49)%;CD34+细胞比例为分别为:(1.25±0.16)%、(9.19±2.10)%;血岛样集落数量分别为0.9±0.36、10.6±0.63;上述结果均以骨髓基质细胞联合细胞因子诱导方法效率最高。基因芯片(hBMSCs/hFLSCs)中总共有240条基因在骨髓基质细胞中高表达,397条基因的在肝脏基质细胞中高表达。对细胞因子、细胞黏附分子以及细胞外基质蛋白的基因加以分析,结果发现在hBMSCs高表达的相关的21条基因,推测可能与造血分化相关。结论人类胚胎干细胞向造血干细胞分化过程中,涉及到细胞与细胞之间的相互作用,相关的细胞因子、细胞黏附分子、细胞外基质蛋白等可能起重要的作用。Objective To investigate the roles of human fetal liver stromal cells （hFLSCs） and human fetal bone marrow stromal cells （hFBMSCs） in the hematopoietic differentiation of human embryonic stem cells and analyze their gene expression profile changes. Methods The embryonic bodies on day 4 were cocultured with hFLSCs or hFBMSC in the presence of cytokines. Flow cytometry was performed after 8 days of induction to detect the expressions of the hemangioblast markers KDR and CD34, and the differential gene expression profiles between hFBMSC and hFLSCs were examined by cDNA microarray analysis. Results Eight days after the induction, （1.06±0.20）% of the hFLSCs and （8.8±1.49）% of the hFBMSCs were positive for KDR, with the positivity rates for CD34 of （1.25 ±0.16）% and （9.17 ±2.10）%, respectively. In hFLSCs and hFBMSCs cultures, 0.9±0.36 and 10.6±0.63 hemagioblast-like cell colonies were found, respectively, cDNA microarray analysis showed that 240 genes were highly expressed in hFBMSCs, and 21 genes related to secreted cytokines, cell adhesion molecules and extracellular matrix proteins were highly expressed. Conclusion The microenvironment including the cell matrix protein and cytokines secreted by the hFBMSCs might play an important role in hemangioblastic differentiation of human bone marrow stromal cells in vitro.

关 键 词：胚胎干细胞 造血干细胞 基因表达 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]