血管内皮生长因子促进小鼠胚胎干细胞的造血分化  

Vascular endothelial growth factor promotes hematopoietic differentiation from mouse embryonic stem cells

作　　者：李府[1] 沈柏均[2] 刘星霞[2] 郑立波[3] 侯怀水[2] 时庆[2] 马秀峰[2] 

机构地区：[1]山东大学第二医院小儿内科,山东济南250033 [2]山东大学齐鲁医院低温医学研究室,山东济南250012 [3]胜利石油管理局胜利医院,山东东营257055

出　　处：《中国病理生理杂志》2006年第8期1602-1605,共4页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助(No.30271248)

摘　　要：目的:研究血管内皮生长因子(VEGF)体外促进小鼠胚胎干细胞系ES-D3向造血分化的能力。方法:先将ES-D3形成拟胚体,将拟胚体细胞转入含不同浓度的VEGF和VEGF+SCF的培养基中。实验分6组,分别为VEGF 5μg/L组、VEGF 10μg/L组、VEGF 20μg/L组、VEGF 5μg/L+SCF组、VEGF 10μg/L+SCF组、VEGF20μg/L+SCF组,同时设不加因子的自发分化对照组。RT-PCR检测造血转录基因GATA-2和早期造血细胞基因c-k it和β-H1的表达,流式细胞仪检测CD34+细胞,甲基纤维素半固体培养法检测生成造血集落的能力。结果:经过1周的诱导培养,实验组生成的细胞可以表达GATA-2、c-k it和β-H1,CD34+细胞的比例也升高,并可形成造血祖细胞的集落。从诱导生成CD34+细胞的比例和生成的集落数量看,VEGF联合SCF组的诱导效率要高于VEGF单用组和对照组,其中以VEGF 20μg/L+SCF组和VEGF 10μg/L+SCF组的诱导效率最高。结论:VEGF能够促进ESC的早期造血分化,尤以与SCF合用时,其诱导效率更高。AIM： To study the effect of vascular endothelial growth factor （VEGF） on hematopoietic differentiation from mouse embryonic stem cells （ ESC ） in vitro. METHODS ： ES - D3 was allowed to grow on mouse fetal fibroblast feeder layer, and then was developed into embryoid bodies （EB）. EB cells were transferred into medium supplemented with different concentration of VEGF and VEGF ＋ SCF for 1 week. Six groups, including. VEGF 5 μg/L, VEGF 10 μg/L, VEGF 20 μg/L, VEGF 5 μg/L ＋ SCF, VEGF 10 μg/L ＋ SCF and VEGF 20 μg/L ＋ SCF, were designed. The group of spontaneous differentiation without cytokine（s） was used as control. Hematopoietic transcription factor GATA -2 and early hematopoietic differentiation genes （ c - kit and β - H1 ） were detected by RT - PCR. The content of CD34 ^＋ ceils in each group were measured by flow cytometry. The cells derived from ESC were incubated in semisolid methycellulose cultures. The numbers of total colony - forming units in culture （ CFU - C） were counted by reverse microscope. RESULTS ： ES - D3 grew and formed EB at day 4. VEGF had a stimulatory effect as a single factor on the expression of genes associated with early hematopoietic differentiation （ GATA - 2, c - kit and β - H1 ）, the generation of CD34 ^＋ cells and CFU - C in EB. The effects of VEGF ＋ SCF were the most potent in the experimental groups according to the percent of CD34 ^＋ cells and the numbers of hematopoietic colonies. The most highest inducing efficacy was achieved in VEGF 20 μg/L or VEGF 10 μg/L combined with SCF. CONCLUSION： VEGF promotes the differentiation of ESC into hematopoietic cells in vitro. The strongest effect was achieved when VEGF was combined with SCF.

关 键 词：胚胎干细胞 内皮生长因子 拟胚体 分化 

分 类 号：R363[医药卫生—病理学]