ORIENTATIONAL DEVELOPMENT AND DIFFERENTIATION OF MOUSE PRIMORDIAL GERM CELLS INTO HEPATOCYTES IN VITRO
Author(s): LIU Xiao-ping, GAO Ying-mao, BING Lu-jun
Pages: 650-654
Year: 2005 Issue: 6
Journal: ACTA ANATOMICA SINICA
Keyword: 原始生殖细胞; 肝细胞; β-神经生长因子; 碱性成纤维细胞生长因子; 表皮生长因子; 维甲酸; 诱导分化; 小鼠;
Abstract: 目的探索诱导小鼠原始生殖细胞向肝细胞定向分化的最佳条件.方法分离提取小鼠的原始生殖细胞,体外培养扩增后,接种到6孔培养板中.向不同孔中分别加入不同浓度的bFGF、EGF、β-NGF、RA、肝细胞提取液和与胎肝组织分隔培养,进行诱导分化.用靛青绿摄入试验和α-1-抗胰蛋白酶(AAT)、白蛋白(ALB)免疫细胞化学染色鉴定原始生殖细胞向肝细胞分化的状况.结果与胎肝组织分隔培养2 d,可见原始生殖细胞向肝样细胞分化,分化细胞呈圆形和星形,圆形细胞多呈簇状,可以摄入靛青绿呈绿色,并呈AAT和ALB免疫阳性着色,培养12 d时分化率达80%.肝细胞提取液也可诱导原始生殖细胞向肝样细胞分化,12 d时分化率达70%以上;0.5g/L和1 g/L的肝细胞提取液的诱导分化率无显著差异;但是0.5 g/L提取液诱导组圆形细胞较多;而1 g/L肝提取液诱导组,星状分化细胞多.β-神经生长因子(β-NGF)诱导4 d,可见肝样细胞出现,随诱导时间延长,分化细胞增多,圆形分化细胞较多,12 d时分化率达60%以上;20 μg/L和50 μg/L两种浓度的β-NGF诱导分化率无显著差异.bFGF、EGF、RA诱导组未见ALB免疫反应阳性细胞.RA可以增强β-NGF的诱导分化作用,使星状细胞增多明显.结论β-NGF和肝细胞因子可诱导原始生殖细胞向肝细胞定向分化.
Pages: 650-654
Year: 2005 Issue: 6
Journal: ACTA ANATOMICA SINICA
Keyword: 原始生殖细胞; 肝细胞; β-神经生长因子; 碱性成纤维细胞生长因子; 表皮生长因子; 维甲酸; 诱导分化; 小鼠;
Abstract: 目的探索诱导小鼠原始生殖细胞向肝细胞定向分化的最佳条件.方法分离提取小鼠的原始生殖细胞,体外培养扩增后,接种到6孔培养板中.向不同孔中分别加入不同浓度的bFGF、EGF、β-NGF、RA、肝细胞提取液和与胎肝组织分隔培养,进行诱导分化.用靛青绿摄入试验和α-1-抗胰蛋白酶(AAT)、白蛋白(ALB)免疫细胞化学染色鉴定原始生殖细胞向肝细胞分化的状况.结果与胎肝组织分隔培养2 d,可见原始生殖细胞向肝样细胞分化,分化细胞呈圆形和星形,圆形细胞多呈簇状,可以摄入靛青绿呈绿色,并呈AAT和ALB免疫阳性着色,培养12 d时分化率达80%.肝细胞提取液也可诱导原始生殖细胞向肝样细胞分化,12 d时分化率达70%以上;0.5g/L和1 g/L的肝细胞提取液的诱导分化率无显著差异;但是0.5 g/L提取液诱导组圆形细胞较多;而1 g/L肝提取液诱导组,星状分化细胞多.β-神经生长因子(β-NGF)诱导4 d,可见肝样细胞出现,随诱导时间延长,分化细胞增多,圆形分化细胞较多,12 d时分化率达60%以上;20 μg/L和50 μg/L两种浓度的β-NGF诱导分化率无显著差异.bFGF、EGF、RA诱导组未见ALB免疫反应阳性细胞.RA可以增强β-NGF的诱导分化作用,使星状细胞增多明显.结论β-NGF和肝细胞因子可诱导原始生殖细胞向肝细胞定向分化.
Citations
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