HGF和FGF4体外诱导人骨髓CD45^-CD117^-细胞向肝细胞分化的研究  

Differentiation of adult bone marrow CD45-CD117'stem cells into hepatocyte in vitro

作　　者：李文晰[1] 段芳龄[1] 马军[1] 陈香宇[1] 

机构地区：[1]郑州大学消化疾病研究所,河南省郑州市450003

出　　处：《世界华人消化杂志》2004年第3期697-701,共5页World Chinese Journal of Digestology

基　　金：河南省卫生厅创新人才基金资助项目;No.2002223~~

摘　　要：目的:探讨在HGF、FGF4诱导下人骨髓CD45-CD117-细胞能否向肝细胞分化,并探讨两种生长因子在诱导分化中可能的作用机制. 方法:骨髓来自4名4-40岁健康志愿者的胸骨或髂骨.用两步磁式分离法分离出CD45-CD117-细胞,分别用含有FGF4,HGF,HGF+FGF4或不加生长因子的DMEM培养基进行培养,相应地分为A,B,C,D四组.分别于新分离时和培养7 d,14 d,21 d,28 d时留细胞.备作免疫细胞化学检测AFP,CK18,白蛋白,PAS染色检测糖原等肝系细胞的特征型标志,RT-PCR检测C、D组细胞c-met(HGF受体)和FGFR2(FGF4R)mRNA表达情况. 结果:A,B,C三组加生长因子诱导培养后的细胞均可检测到肝系细胞的标志,D组细胞不加生长因子培养后没有检测到肝系细胞的标志.c-met和FGFR2mRNA在新分离的和D组培养7 d,14 d时的细胞均有较弱的表达,而在C组细胞诱导培养7 d,14 d时表达升高. 结论:HGF和FGF4可诱导人骨髓CD45-CD117-细胞向肝细胞样细胞分化.生长因子诱导分化的作用可能通过与其受体之间的正反馈调节.AIM: To observe whether the CD117-CD45 bone marrow cell can differentiate into the cells with characters of hepa-tocyte by the stimulation of hepatocyte growth factor (HGF) and fibroblast growth factor-4 (FGF4), and to explore the acting mechanism of the growth factors. METHODS: Bone marrow cells were from the stern or ilium of 4 healthy volunteers between 4-40 years old. CD117 CD45 bone marrow cells separated by magnetic cell sorting method, cultured in DMEM medium with FGF4, HGF, FGF4+ HGF or no growth factor were divided into four groups: groups A, B, C, and D accordingly. Cells were collected on days 0, 7, 14, 21, and 28 for detecting the hepatocyte markers: AFP, CK18, albumin by immunocytochemistry, glycogen by PAS staining and c-met, FGFR2 mRNA by RT-PCR. RESULTS: The specific markers of hepatocyte were positive in groups A, B and C but not in group D. C-met and FGFR2 mRNA expressed at a low level in the cells newly isolated or cultured in the medium without growth factor, but at increasing level after cutured with growth factor. CONCLUSION: HGF and FGF4 can induce CD45-CD117 differentiate into hepatocyte-like cells. The effect of growth factor on cell differentiation is probably by the positive regulation between the growth factor and its receptor.

关 键 词：HGF FGF4 体外诱导 骨髓干细胞 CD45^- 细胞 CD117^-细胞 肝细胞 细胞分化 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]