骨髓内皮细胞条件培养液促进小鼠胚胎干细胞向造血分化  

Bone Marrow Endothelial Cell-Conditioned Medium Promotes Hematopoietic Differentiation of Mouse Embryonic Stem Cells

作　　者：赵惠萍[1] 卢光琇 王绮如[1] 

机构地区：[1]中南大学湘雅医学院血液生理研究室,长沙410078 [2]湖南省干细胞工程技术研究中心,长沙410078

出　　处：《中国实验血液学杂志》2003年第2期109-114,共6页Journal of Experimental Hematology

基　　金：国家自然科学基金重点项目资助编号 3 0 0 3 0 0 70

摘　　要：为观察骨髓内皮细胞条件培养液 (BECM)和 (或 )细胞因子 (VEGF +SCF +EPO)对小鼠胚胎干细胞 (ESC D3细胞 )生成造血干 /祖细胞的促进作用 ,先将ESC D3细胞形成 4天胚状体 (Day 4embryoidbody,4dEB) ,再诱导 4dEBs生成造血干 /祖细胞。实验分 4组 ,第 1,2和 3组分别为BECM +VEGF +SCF +EPO ,BECM和VEGF +SCF +EPO组 ,第 4组为自发分化对照组。检测各组造血干 /祖细胞特异抗原、造血转录因子表达以及造血集落的形成。结果显示 ,BECM和 (或 )细胞因子诱导生成的细胞均表达造血干 /祖细胞抗原 (c kit,Sca 1,Thy 1和CD34)和造血转录因子(c myb ,SCL和β H1) 基因mRNA ,培养后可产生HPP CFC和BFU E。从诱导ESC D3细胞生成造血干 /祖细胞的数量和生成的集落总数看 ,BECM联合细胞因子组诱导效率均显著高于单用组和对照组。结论 :骨髓内皮细胞条件培养液能显著促进胚胎干细胞早期造血分化 。The purpose of this study was to observe the bone marrow endothelial cell conditioned medium (BECM) and cytokines, i.e. vascular endothelial growth factor (VEGF), stem cell factor (SCF) and EPO promoting the generation of hematopoietic precursor cells from mouse embryonic stem cells (ESC) in vitro. Day 4 embryoid body (4dEB) cells were derived from ESC D3 cell line, a murine ESC line, and then induced with BECM and/or cytokines. Four groups, i.e. BECM, BECM+VEGF+SCF+EPO, VEGF+SCF+EPO and control (spontaneous differentiation), were designed. Immunochemistry staining and flow cytometry were adopted to observe the antigen expression, RT PCR to detect hematopoietic transcription factors, and hematopoietic progenitor assay to examine hematopoietic differentiation. The results showed that the cells induced from ESC expressed hematopoietic precursor cell antigens(c kit, Sca 1, Thy 1 and CD34), transcription factors(c myb, SCL and β H1) and generated HPP CFC and BFU E. The effect of BECM+VEGF+SCF+EPO was the most potent in the inducing groups according to the numbers of hematopoietic precursor cells and colonies. It is concluded that BECM promotes the differentiation of ESC into hematopoietic precursor cells in vitro, and this effect is the strongest when BECM combining with VEGF+SCF+EPO.

关 键 词：骨髓内皮细胞 培养液 细胞因子 胚胎干细胞 造血干细胞 胚状体 骨髓移植 基因治疗 

分 类 号：R457.7[医药卫生—治疗学] R331.22[医药卫生—临床医学]