摘要
目的 初步探讨了酸性成纤维细胞生长因子 (aFGF)及肝细胞生长因子 (HGF)对大鼠胎肝干细胞分化成熟的影响。方法 采用胶原酶消化法分离胎龄 13d的SD大鼠胎肝细胞 ,分别接种于Ⅰ、Ⅱ、Ⅲ、Ⅳ组细胞培养瓶中进行体外培养。Ⅰ组 :培养液中含酸性成纤维细胞生长因子(aFGF)和肝细胞生长因子 (HGF) ;Ⅱ组 :培养液中含aFGF ;Ⅲ组 :培养液中含HGF ;Ⅳ组培养液中无细胞因子 ,此组作为对照组。检测在相同培养时间 (第 3、5、7、9、11天 )细胞培养上清液中甲胎蛋白(AFP)和白蛋白 (ALB)的含量 ;并用免疫细胞化学法鉴定在无细胞因子干预培养 3d后 ,胎肝干细胞对细胞角质素 19(CK 19)的表达。结果 Ⅰ组、Ⅱ组及Ⅲ组分别与Ⅳ组比较 ,在相同培养时间的培养上清液中AFP的含量均较少 ,而ALB的含量均较多 ,且差异均有显著性意义 (AFP :t=6 2 6 4 ,3 2 2 1,2 5 91,P <0 0 5 ;ALB :t=6 32 4 ,2 14 5 ,2 95 4 ,P <0 0 5 ) ;Ⅱ组和Ⅲ组相同培养时间的培养上清液中AFP及ALB的含量差异均无显著性意义 (AFP :t =0 5 0 1,P >0 0 5 ;ALB :t =0 6 16 ,P >0 0 5 ) ;而Ⅳ组分别与Ⅱ组和Ⅲ组比较 ,在相同培养时间的培养上清液中ALB的含量均较少 ,而AFP的含量均较多 ,差异也均有显著性意义 (AFP :t =3 2 2 1,2 5 91,P
Objective To evaluate the effect of acidic fibroblast growth factor (aFGF) and hepatocyte growth factor (HGF) on differentiation and maturation of murine fetal hepatic stem cells. Methods ED13 Sprague-Dawley (SD) fetal rat liver cells were isolated by collagenase digestion technique and cultured under different conditions (groupⅠ,Ⅱ, Ⅲ, and Ⅳ).aFGF and HGF were used in group Ⅰ, aFGF in group Ⅱ, HGF in group Ⅲ, and group Ⅳ was used as the control.The AFP and ALB in culture matrix were determined by radioimmunoassay (RIA) and the expression of CK-19 was observed by immunocytochemical technique. Results The values of AFP in group Ⅰ, Ⅱ, and Ⅲ were lower than those in the control group (group Ⅳ), while ALB were higher (AFP: t=6.264, 3.221, 2.591, P<0.05;ALB: t=6.324, 2.145, 2.954, P<0.05).AFP and ALB in group Ⅱ and Ⅲ showed no significant difference (AFP: t=0.501, P>0.05;ALB: t=0.616, P>0.05).AFP in group Ⅳ was higher than that in group Ⅱor Ⅲ, but ALB was lower (AFP: t=3.221, 2.591, P<0.05;ALB: t=2.145, 2.954, P<0.05).The CK-19 + cells were over 50% in all samples, and they were over 90% in 22 samples (73.33%). Conclusion It is feasible to isolate the hepatic stem cells from ED13 fetal rat liver by collagenase digestion technique.It is proved that both aFGF and HGF can precipitate hepatic stem cells to differentiate into mature hepatocytes, and there is synergistic action between them.
出处
《中华放射学杂志》
CAS
CSCD
北大核心
2004年第2期124-128,共5页
Chinese Journal of Radiology
基金
高等学校特聘教授匹配骨干教师项目 (苏教人 [2 0 0 1] 45号 )
关键词
酸性成纤维细胞生长因子
肝细胞生长因子
肝干细胞
胶原酶消化法
Rats, Sprague-Dawley
Liver, stem cell
Cell cultivation
Fibroblast growth factor,acidic
Hepatocyte growth factor
Cytokeratin
