不同浓度HGF、EGF优化大鼠胎肝干细胞体外培养的研究  

Different Concentrations of Hepatocyte Growth Factor and Epidermal Growth Factor in the Regulation of Fetal Liver Stem Cells in Rats

作　　者：赵戈[1] 龚振斌[2] 黄启科[2] 党立力[2] 张宁[2] 王廷[1] 吕勇刚[1] 陶开山[1] 

机构地区：[1]第四军医大学附属西京医院血管内分泌科,陕西西安710032 [2]第四军医大学附属西京医院肝胆胰脾外科,陕西西安710032

出　　处：《现代生物医学进展》2013年第9期1622-1626,共5页Progress in Modern Biomedicine

基　　金：国家自然科学基金项目(81170419;30772094;81101619)

摘　　要：目的：研究不同浓度肝细胞生长因子（Hepatocyte growth factor,HGF）和表皮生长因子（epidermal growth factor,EGF）对大鼠胎肝干细胞体外增殖的影响,探索二者间有无协同作用。方法：取孕14天胎龄F344大鼠胚胎的胎肝,经三步分离法分离纯化后,配置不同浓度HGF、EGF及HGF和EGF联合组培养基,将胎肝干细胞分组培养。光镜下观察细胞增殖状况,MTT法观察不同浓度和不同时间HGF、EGF及HGF和EGF联合对大鼠胎肝干细胞增殖的影响,并进行统计学分析。结果：HGF 10~80 ng/mL各浓度组,EGF 10~80 ng/mL各浓度组增殖效应均大于对照组。当HGF为20 ng/mL,增殖效应明显大于对照组（P〈0.01）,当HGF浓度继续增高时,增殖效应无明显增高。将20 ng/mL HGF组与不同浓度EGF组合后分组培养细胞,发现20 ng/ml HGF和10 ng/mLEGF联合组增殖效应明显升高,继续升高联合组中EGF浓度,增殖效应无明显提高。结论：HGF和EGF具有明显改善大鼠胎肝干细胞体外无血清培养条件的作用,二者对大鼠胎肝干细胞体外培养具有协同促进作用。其中20 ng/mL HGF和10 ng/mL EGF联合培养促进大鼠胎肝干细胞增殖效果最明显。Objective： To research the proliferation effect of HGF and EGF to the rat fetal liver stem cells at different concentration in vitro. Methods： Using Three-step separation method, fetal liver stem cells were purificated from embryonic liver tissue of pregnant 14 days F344 rats. Cells were treated with condition medium which contains different concentration of HGF and EGF. Cellular proliferation state was observed by electron microscopy. Cellular proliferation of different culture medium was determined by MTT assay. Results： The proliferation effect of HGF 10 - 80 ng/ml in each concentration group and EGF 10 - 80 ng/ml in each concentration group is greater than that in the control group. As HGF for 20 ng/mL, proliferation effect was significantly greater than control group （P〈0.01）. As HGF c- oncentration continues increasing, no obvious increase of proliferation effect was observed. In 20 ng/mL HGF group combined with different concentration EGF group, we found that in 20 ng/ml HGF withl0 ng/mL EGF combination groups, the proliferation effect was increased significantly. As the EGF concentration continues to risen in joint group, the proliferation effect was not improved obviously. Cunelusion： HGF and EGF play an important role in improving the serum-free culture conditions of rat fetal liver stem cells. And they have collaborative stimulative effect on rat fetal liver stem cells proliferation in vitro. 20 ng/ml HGF combined with 10 ng/mL EGF has significant effect on cellular proliferation of rat fetal liver stem cells.

关 键 词：胎肝干细胞 肝细胞生长因子 表皮生长因子 细胞增殖 

分 类 号：Q95-3[生物学—动物学] Q813