Organogenesis of established ES cells and its application for clinical medicine.

已建立的ES细胞的器官发生及其在临床医学中的应用。

• 批准号: 12470247
• 负责人: ISHIKAWA Hiroshi
• 金额: $ 8.83万
• 依托单位: JIKEI UNIVERSITY SCHOOL OF MEDICINE
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12470247/
• 关键词: Hepatocyte; ES Cell; embryotrophic factor; cellular transplantation; Bio-hybrid typed liver apparatus; spontaneous dwarf rat; primordial liver; albumin; 肝細胞移植医療; バイオ人工肝臓; ES細胞; アルブミン; プロトロビン; IGF-1; 胆汁産生; urea cycle; マウスEES細胞; ラットEES細胞

A cell line with the characteristics of hepatocytes was established from rat early embryonic stem cells (REEES). This cell line was established using a new novel method of us et al. from two cell embryos taken from the spontaneous dwarf rat (SDR). The hepatocyte cell line (REES-hep) was instituted from dark red colored tissue in embryos during embryogenesis using REES cell line cultured in the presence of embryotrophic factors. These cell lines were cultured with DMEM/F12 medium supplemented 10% FBS and 1ng/ml of LIF. They were found to maintain their diploid state, were characterized with 42 normal chromosomed and proliferated to confluence, contact inhibition was also present. These cells produced albumin when cultured using a collagen sponge gel system and reconstructed in a funicular form resembling the cell cords of liver. The cells also produced albumin and bilirubin when transplanted into the spleen of SDR. Also, transplantation of REES-hep cells into the spleen of liver-insufficient rat promoted prolongation of its life. Reconstruction of a REES-hep cell line from early embryonic stem cells should help in treating hepatic insufficient patients. It will be valuable for further research, as an introduction to cell transplantation and application for use in a bio-hybrid typed liver apparatus.

从大鼠早期胚胎干细胞（REEES）中建立了具有肝细胞特性的细胞系。该细胞系是采用美国等的一种新方法，从自发性侏儒大鼠（SDR）的两细胞胚胎中建立的。肝细胞系（REES-hep）是在胚胎发生过程中，使用在胚胎营养因子存在下培养的REES细胞系，从胚胎中的暗红色组织中建立的。这些细胞系用补充10%FBS和lng/ml LIF的DMEM/F12培养基培养。结果表明，它们保持二倍体状态，具有42条正常染色体，并增殖至融合，同时存在接触抑制。当使用胶原海绵凝胶系统培养时，这些细胞产生白蛋白，并以类似于肝细胞索的索状形式重建。当移植到SDR的脾脏中时，细胞还产生白蛋白和胆红素。此外，REES-hep细胞移植到肝功能不全大鼠的脾脏中促进其寿命的延长。从早期胚胎干细胞重建REES-hep细胞系有助于治疗肝功能不足的患者。为细胞移植及在生物混合型肝脏装置中的应用提供了参考。

项目成果

Ishiwata I: "Effects of embryotrophic factor on embryogenesis of mouse embryos"Proceedings of the 17^<th> world congress of IFFS 2001. 2001. 133-137 (2001)

Ishiwata I：“胚胎营养因子对小鼠胚胎胚胎发生的影响”IFFS第17届世界大会记录2001. 2001. 133-137 (2001)

Ishiwata I et al.: "Properties of mouse embryonic stem cells established by a novel method"Proceedings of the 17^<th> world congress of IFFS 2001. 2001. 139-145 (2001)

Ishiwata I等人：“通过新方法建立的小鼠胚胎干细胞的特性”IFFS第17届世界大会记录2001.2001.139-145(2001)

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Tokieda Y：“建立一种冷冻保存和解冻小鼠卵巢的新方法”《人类细胞》。

Horikawa R: "Regulation of pituitary growth hormone-secretagogue and growth Hormone-releasing homone receptor RNA Expression in young dwarf rat"Endocrine Journal. 47(Suppl). S53-S56 (2000)

Horikawa R：“幼年侏儒大鼠垂体生长激素促分泌剂和生长激素释放激素受体 RNA 表达的调节”内分泌杂志。

Kiguchi K: "Establishment and characterization of a lymphoepithelial-like carcinoma cell line (HUUCLEC) derived from the human uterine cervix"Human Cell. 15(2). 97-102 (2002)

Kiguchi K：“源自人子宫颈的淋巴上皮样癌细胞系 (HUUCLEC) 的建立和表征”Human Cell。