Development of efficient process for isolation, propagation and differentiation induction of normal human hepatic stem cells and its application to artificial liver

正常人肝干细胞高效分离、增殖和分化诱导工艺的开发及其在人工肝中的应用

• 批准号: 15560681
• 负责人: MATSUSHITA Taku
• 金额: $ 2.37万
• 依托单位: Sojo University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15560681/
• 关键词: hepatic stem cells; hepatoblast; artificial liver; regenerative medicine; human fetal hepatocytes; sodium butyrate; PIVKA-II; transformation

1, Studies on transformation of normal human hepatic stem cells in propagation process and its preventionIf the normal human hepatic stem cells (hepatoblast) transform in propagation process, these cells can not be applicable to regenerative medicine and artificial liver. In fact, the stem cells and transformed tumor cells have similar characteristics and makers. In this research, we developed the examination method for the transformation of hepatic stem cells (hepatoblast) using PIVKA-II (abnormal prothrombin) as a maker, which is clinical tumor maker of hepatocytes. We indicated that human fetal hepatocytes including hepatoblasts started to transform at the8th passage of propagation by using PIVKA-II assay. And we also indicated that induction of hepatoblast from human fetal hepatocytes teated with Sodium butyrate, caused the transformation depended on the concentration of sodium butyrate. It was also found that Vitamin K has prevention effect of the transformation of human fetal heptaocytes.2,Studies on the materials of culture substrate for high density culture of hepatoblast.For development of artificial liver module using human hepatoblast, it is necessary to study on material characteristics of culture substrate for high density culture, functional expression and prevention of transformation of human hepatoblast. In this research, we examined several culture substrates. Macroporous siran material was better for high density culture. And porous hydroxyapatite was better for cytochrome P450 activity expression of hepatoblast.

1、正常人肝干细胞在增殖过程中的转化及其预防研究正常人肝干细胞（成肝细胞）在增殖过程中如果发生转化，就不能应用于再生医学和人工肝。事实上，干细胞和转化的肿瘤细胞具有相似的特征和标志物。在本研究中，我们开发了使用PIVKA-II（异常凝血酶原）作为标记物（肝细胞的临床肿瘤标记物）的肝干细胞（成肝细胞）转化的检测方法。我们用PIVKA-II法检测了人胎肝细胞包括肝母细胞在第8代开始转化。丁酸钠诱导人胎肝细胞向肝细胞转化时，其转化程度依赖于丁酸钠的浓度。维生素K对人胎肝细胞转化有一定的预防作用。2、人肝细胞高密度培养基质材料的研究，研究人肝细胞高密度培养基质的材料特性、功能表达及预防转化的作用，为研制人肝细胞人工肝模块奠定基础。在这项研究中，我们研究了几种培养基。大孔材料有利于高密度培养。多孔羟基磷灰石对细胞色素P450活性表达有较好的促进作用。

项目成果

人工肝臓開発のための細胞源の検討-肝幹細胞の利用とがん化(形質転換)の抑制-

人工肝开发的细胞来源的思考 - 肝干细胞的利用和癌变（转化）的抑制 -

• 发表时间: 2004
• 期刊: 化学工学会沖縄大会研究発表講演要旨集
• 作者: Y Matsumoto;Y Kadota;Y Matsumoto;Y Kadota;Y Matsumoto;松下 琢
• 通讯作者: 松下 琢

正常ヒト肝幹細胞を用いた人工肝臓開発のための基礎的研究

利用正常人肝干细胞开发人工肝的基础研究

• 发表时间: 2004
• 期刊: 化学工学会秋田大会研究発表講演要旨集
• 作者: Y Matsumoto;Y Kadota;Y Matsumoto;Y Kadota;Y Matsumoto;松下 琢;松下 琢
• 通讯作者: 松下 琢

T.Matsushita: "Spheroid formation and functional restoration of human fetal hepatocytes on poly-amino acid-coated dishes after serial proliferation"Cytotechnology. 42巻2号. 57-66 (2003)

T.Matsushita：“连续增殖后聚氨基酸包被培养皿上人胎儿肝细胞的球体形成和功能恢复”《细胞技术》第 42 卷，第 2. 57-66 期（2003 年）。

Novel mechanism of hybrid liposomes-induce apoptosis in human tumor cells.

混合脂质体诱导人类肿瘤细胞凋亡的新机制。

• 发表时间: 2005
• 期刊: International Journal of Cancer 115巻(In press)
• 作者: Y Matsumoto;Y Kadota;Y Matsumoto;Y Kadota;Y Matsumoto
• 通讯作者: Y Matsumoto

The basic research for artificial liver development using normal human hepatic stem cells.

利用正常人肝干细胞开发人工肝的基础研究。

• 发表时间: 2004
• 期刊: Proceedings of Akita meeting of chemical engineering society of Japan. SA218
• 作者: Y Matsumoto;Y Kadota;Y Matsumoto;Y Kadota;Y Matsumoto;松下 琢;松下 琢;清田 章文;T Matsushita;T Matsushita
• 通讯作者: T Matsushita