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的研究如果正常人肝干细胞（成肝细胞）在增殖过程中发生转化，则这些细胞不能适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记。本研究开发了以PIVKA-II（异常凝血酶原）为标记物的肝干细胞（成肝细胞）转化的检查方法，PIVKA-II是肝细胞的临床肿瘤标记物。我们通过PIVKA-II检测表明，人胎儿肝细胞包括成肝细胞在增殖第8代时开始转化。我们还指出，用丁酸钠诱导人胎儿肝细胞成肝细胞，引起的转化取决于丁酸钠的浓度。还发现维生素K对人胎儿肝细胞的转化有预防作用。2、肝母细胞高密度培养的培养基质材料的研究。利用人肝母细胞开发人工肝模块，有必要进行研究。人肝细胞高密度培养培养基质特性、功能表达及预防转化。在这项研究中，我们检查了几种培养基质。大孔Siran材料更适合高密度培养。多孔羟基磷灰石对肝细胞色素P450活性表达效果较好。

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

ヒト肝幹細胞の効率的取得とスフェロイド形成による肝機能向上に関する研究

通过高效获取人肝干细胞及球体形成改善肝功能的研究

• 发表时间: 2004
• 期刊: 第41回化学関連支部合同九州大会講演予稿集
• 作者: Y Matsumoto;Y Kadota;Y Matsumoto;Y Kadota;Y Matsumoto;松下 琢;松下 琢;清田 章文
• 通讯作者: 清田 章文

Studies on cell sources for artificial liver development.-use of hepatic stem cells and prevention of transformation of the cells-

人工肝发育细胞来源的研究-肝干细胞的利用及细胞转化的预防-

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

松下 琢: "ヒト肝幹細胞の効率的取得と人工肝臓・再生医療への応用に関する基礎研究"近未来新素材研究会 第3回講演会要旨集. 8-9 (2003)

松下卓：“人类肝脏干细胞的高效获取及其在人工肝和再生医学中的应用的基础研究”近期新材料研究组第3次讲座摘要（2003年）。

松下 琢: "酪酸ナトリウムによるヒト肝芽細胞の誘導に関する研究"再生医療. 3巻増刊号. 79 (2004)

Taku Matsushita：“丁酸钠诱导人成肝细胞的研究”再生医学第3卷特刊（2004年）。