The relationship between differentiation and proliferation in late gestation fetal rat hepatocytes

作     者：Gruppuso, PA Bienieki, TC Faris, RA 

作者机构：Rhode Isl Hosp Div Pediat Endocrinol Providence RI 02903 USA Brown Univ Providence RI 02903 USA 

出 版 物：《PEDIATRIC RESEARCH》 (儿科研究)

年 卷 期：1999年第46卷第1期

页      面：14-19页

核心收录：

学科分类：1002[医学-临床医学] 100202[医学-儿科学] 10[医学] 

基　　金：NCI NIH HHS [CA66005] Funding Source: Medline NICHD NIH HHS [HD11343, HD24455] Funding Source: Medline 

主　　题：动物 新生 生物学标记/分析 细胞黏附分子/分析 细胞分化/药物作用 细胞分裂/药物作用 细胞 培养的 DNA/生物合成 胎儿 孕龄 葡糖激酶/分析 肝细胞生长因子/药理学 肝/细胞学 肝/药物作用 肝/胚胎学 磷酸烯醇丙酮酸羧激酶(GTP)/分析 转化生长因子α/药理学 甲胎蛋白类/分析 动物 大鼠 

摘      要：Hepatocyte proliferation and differentiation occur simultaneously during late mammalian gestation, We hypothesized that regulation of hepatocyte growth and differentiation would be coordinated in late gestation fetal hepatocyte cultures such that proliferation would be most active in a population of less well-differentiated cells. Cultured fetal hepatocytes (embryonic d 19 and 21;E19 and E21) were studied using double staining immunofluorescent microscopy. Differentiation was assessed as staining for cw-fetoprotein (AFP), three markers of enzymic differentiation (glucokinase [GK], phosphoenolpyruvate carboxykinase [PEPCK], and carbamoyl phosphate synthase [CPS]), and a hepatocyte cell-cell adhesion molecule (C-CAM). Proliferation was assessed using immunocytochemical detection of proliferating cell nuclear antigen (PCNA) or 5-bromo-2 -deoxy-uridine (BrdU) incorporation into DNA. Fetal hepatocyte cultures consisted of a heterogeneous population of cells, slightly more than half of which were proliferative under defined, growth factor-free conditions. These cultures were heterogeneous for AFP expression. There was no correlation between the expression of AFP and PCNA or AFP and S-phase entry (BrdU staining) during the first 48 h in culture. Similar results were obtained in staining for the enzymic differentiation markers and C-CAM. In addition, the differentiation status of cultured fetal hepatocytes was unrelated to a presumed indicator of mature growth regulation, mitogenic responsiveness to transforming growth factor alpha (TGF alpha), and hepatocyte growth factor (HGF). Finally, absence of any correlation between proliferation and differentiated phenotype was supported by in vivo studies using staining for PCNA, AFP, CPS, and PEPCK in liver sections. These results indicate that the developmental program governing differentiation of late gestation fetal rat hepatocytes is independent from mechanisms controlling proliferation.