Establishment of a isolation and maturation system for human adult hepatic stem cells

人成体肝干细胞分离成熟体系的建立

• 批准号: 14370384
• 负责人: NAKATSUJI Norio
• 金额: $ 8.9万
• 依托单位: KYOTO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14370384/
• 关键词: human hepatic stem; progenitor cells; mesenchymal cells; specific surface marker genes; differentiation; subcloning

Previously, we have isolated hepatic stem/progenitor cells from adult liver tissue of the transgenic mouse, in which green fluorescent protein was expressed highly in endodermal cells. Then, we have investigated the specific surface marker genes for hepatic stem/progenitor cells. Consequently, we identified several genes for hepatic stem/progenitor cells: In order to determine the specificity of these genes including several EST clones for hepatic stem/progenitor cells, we are now going to verify the difference of gene and also protein product expression between hepatic stem/progenitor cells and mature adult hepatocytes. Futhermore, we are going to investigate the human homologues of these genes subcloned into plasmid after PCR amplification. On the other hand, we confirmed the fact that human hepatic stem/progenitor cells were obtained from human adult liver tissue using the same method for fetal and adult mouse hepatic stem/progenitor cells as we previously reported (Yasuchika K, et al. Hepatology, 2002)(Azuma H, et al. Hepatology, 2003). During further profiling for the specific surface marker expression of mouse hepatic stem/progenitor cells-enriched fraction, we also identified that Thy1-positive mesenchymal cells contained in the mouse fetal liver promoted the maturation of CD49f-positive hepatic stem/progenitor cells in vitro (Hoppo T, et al. Hepatology, 2004, in press). It is anticipated that this kind of mesenchymal cells or recombinant proteins associated with them would be necessary for the maturation of human hepatic stem/progenitor cells into functional adult hepatocytes in vitro. Therefore, we are now executing further investigation for the characteristics of these mesenchymal cells. As described above, we are going to reveal the specific marker genes and also establish the differentiation system for hepatic stem progenitor cells.

在此之前，我们已经从转基因小鼠的成年肝组织中分离出肝干/祖细胞，其中绿色荧光蛋白在内胚层细胞中高度表达。然后，我们研究了肝干/祖细胞的特异性表面标记基因。因此，我们鉴定了肝脏干细胞/祖细胞的几个基因：为了确定这些基因（包括肝脏干细胞/祖细胞的几个EST克隆）的特异性，我们现在将验证肝脏干细胞/祖细胞之间基因和蛋白质产物表达的差异。成熟的成年肝细胞。接下来，我们将对这些基因的同源基因进行研究。另一方面，我们证实了这样的事实，即人肝干/祖细胞是使用与我们先前报道的胎儿和成年小鼠肝干/祖细胞相同的方法从人成年肝组织中获得的（Yasuchika K，et al. Hepatology，2002）（Azuma H，et al. Hepatology，2003）。在进一步分析小鼠肝干/祖细胞富集部分的特异性表面标志物表达的过程中，我们还鉴定了小鼠胎肝中含有的Thy 1阳性间充质细胞促进了CD 49 f阳性肝干/祖细胞的体外成熟（Hoppo T，et al. Hepatology，2004，in press）。这种间充质细胞或与之相关的重组蛋白是体外诱导人肝干/祖细胞成熟为功能性成体肝细胞所必需的。因此，我们现在正在对这些间充质细胞的特性进行进一步的研究。如上所述，我们将揭示特异性标记基因，并建立肝干/祖细胞的分化系统。

项目成果

Yasuchika K et al.: "Establishment of a highly efficient gene transfer system for mouse fetal hepatic progenitor cells."Hepatology. 36(6). 1488-1497 (2002)

Yasuchika K 等人：“建立小鼠胎儿肝祖细胞高效基因转移系统。”肝病学。

Yasuchika K, Hirose T, et al.: "Establishmen of efficient gene transfer system for mouse fetal hepatic progenitor cells"Hepatology. 6. 534-543 (2002)

Yasuchika K、Hirose T 等人：“小鼠胎儿肝祖细胞有效基因转移系统的建立”肝病学。

Azuma H, Hirose T, Fujii H, Oe S, Yasuchika K, Fuj ikawa T, Yamaoka Y.: "Enrichment of hepatic progenitor cells from adult mouse liver."Hepatology.. 37(6). 1385-1394 (2003)

Azuma H、Hirose T、Fujii H、Oe S、Yasuchika K、Fuji ikawa T、Yamaoka Y.：“从成年小鼠肝脏中富集肝祖细胞。”Hepatology.. 37(6)。

Yasuchika K, Hirose T, Fujii H, Oe S, Hasegawa K, Fujikawa T, Azuma H, Yamaoka Y.: "Establishment of a highly efficient gene transfer system for mouse fetal hepatic progenitor cells."Hepatology. 36(6). 1488-1497 (2002)

Yasuchika K、Hirose T、Fujii H、Oe S、Hasekawa K、Fujikawa T、Azuma H、Yamaoka Y.：“建立小鼠胎儿肝祖细胞高效基因转移系统。”肝病学。

Azuma H, Hirose T, et al.: "Hepatic progenitor cells from adult mouse liver"Hepatology. (in press). (2003)

Azuma H、Hirose T 等人：“来自成年小鼠肝脏的肝祖细胞”肝病学。