Development of a hybrid artificial liver with hepatocytes derived embryonic stem cell for chronic liver failure patients

为慢性肝衰竭患者开发具有肝细胞来源的胚胎干细胞的混合人工肝

• 批准号: 16206079
• 负责人: KAJIWARA Toshihisa
• 金额: $ 31.95万
• 依托单位: KYUSHU UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 财政年份: 2004
• 资助国家: 日本
• 起止时间: 2004 至 2005
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-16206079/
• 关键词: embryonic stem cell; organoid; differentiation; hepatocyte; hollow fiber; hybrid artificial liver; regenerative medicine

We investigated the use of organoid culture technique to mediate hepatocyte differentiation of murine and primate embryonic stem cell. We used our original organoid culture technique, which induce cultured cells to form an organoid in lumen of hollow fibers by centrifugal force.Mouse ES cells formed an organoid inside of hollow fibers. The ES cell proliferated for more than 20 days of culture and high density culture was achieved. The expressions of mRNAs of hepatocyte specific genes were detected at 15 days of culture, and were sustained for more than 30 days of culture. The functions of albumin secretion and ammonia removal were evaluated under hepatocyte differentiation culture condition. The level of these functions of the ES cells per unit volume of cell culture space increased as culture time, and reached the same level of primary mouse hepatocytes.Cynomolgus monkey ES cell was also investigated as a model of primate ES cell. Cynomolgus monkey ES cells formed an organoid and expressed some liver-specific functions such as albumin secretion and ammonia removal. The activity of ammonia removal of ES cells organoid per unit volume of cell culture space was roughly comparable to that of primary hepatocytes and human hepatoblastoma cell lines.In conclusion, our developed novel organoid culture technique directed both mouse and cynomolgus monkey ES cells along a hepatocyte lineage. This culture technique seems to be a promising technique to develop a hybrid artificial liver.

我们研究了类器官培养技术在诱导小鼠和灵长类胚胎干细胞向肝细胞分化方面的应用。我们使用了我们独创的类器官培养技术，通过离心力诱导培养的细胞在中空纤维的管腔内形成类器官，小鼠的ES细胞在中空纤维内形成类器官。ES细胞经20天以上的培养扩增，达到高密度培养。培养15d即可检测到肝细胞特异性基因mRNAs的表达，并持续培养30d以上。在肝细胞分化培养条件下评价白蛋白分泌和氨排出功能。单位体积培养空间内ES细胞的上述功能水平随着培养时间的延长而增加，达到原代小鼠肝细胞的水平。食蟹猴ES细胞也作为灵长类ES细胞的模型进行了研究。食蟹猴ES细胞形成类器官，并表达一些肝脏特有的功能，如白蛋白分泌和氨的清除。单位细胞培养空间的ES细胞类有机物的氨去除活性与原代肝细胞和人肝母细胞瘤细胞系大致相当。综上所述，我们开发的新型类有机物培养技术使小鼠和食蟹猴ES细胞沿着一个肝细胞谱系发展。这种培养技术似乎是开发混合型人工肝的一种很有前途的技术。

项目成果

最新医療を支える成形加工技術の現状

支持最新医疗的成型技术现状

• 发表时间: 2005
• 期刊: プラスチックス 56
• 作者: S.Uchida;N.Mizuno;T. Sakiyama;中澤浩二 他
• 通讯作者: 中澤浩二 他

繊維便覧(第4部門4・11・1節 人工肝臓)

纺织手册（第 4 部分第 4、11、1 人工肝）

• 发表时间: 2004
• 作者: Shigeru Naito;Munehiko Minoura;Munenobu Takeda;H. YOSHINO;福田淳二 他
• 通讯作者: 福田淳二 他

人工臓器開発における化学工学の役割

化学工程在人工器官开发中的作用

• 发表时间: 2005
• 期刊: ケミカルエンジニヤリング 50
• 作者: T.Higuchi;S.Yamaguchi;他3名;H. YOSHINO;水本博 他
• 通讯作者: 水本博 他

PUF/スフェロイド型人工肝臓モジュール内でのES細胞の肝細胞への分化誘導

PUF/球体型人工肝模块中ES细胞向肝细胞的诱导分化

• 发表时间: 2005
• 期刊: 人工臓器 34
• 作者: F. Shimojo;S. Kodiyalam;I. Ebbsjo;R. K. Kalia;et. al.;松本欣也 他
• 通讯作者: 松本欣也 他

Establishment of a novel hepatocyte organoid culture and its application to a hybrid artificial liver

新型肝细胞类器官培养物的建立及其在混合人工肝中的应用

• 发表时间: 2004
• 期刊: ABSTRACT BOOK of the Joint Meeting of the Tissue Engineering Society International and the European Tissue Engineering society
• 作者: Tomonori KAWABATA;Masaki KATO;Tomoo MIZUGAKI;Kohki EBITANI;Kiyotomi KANEDA;H.Mizumoto et al.
• 通讯作者: H.Mizumoto et al.