Development of regeneration, purification, and transplantation of cardiomyocytes from mouse and common marmoset ES cells

小鼠和普通狨猴 ES 细胞的心肌细胞再生、纯化和移植的开发

• 批准号: 17209029
• 负责人: FUKUDA Keiichi
• 金额: $ 31.7万
• 依托单位: Keio University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 财政年份: 2005
• 资助国家: 日本
• 起止时间: 2005 至 2007
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-17209029/
• 关键词: human embryonic stem cell; common marmosset; Regeneratie Medicine; Cardiomyocyte; differentiation; Development; Noggin; Cell Transplantation; 胚性幹細胞; 奇形腫; 多分化能; 転写因子; NKX2.5; イオンチャネル; 心筋梗塞

This Study tried to establish a novel method for generation, purification, and transplantation of new cardiomyocytes from human and common marmoset ES cells. We found that Noggin induce differentiation from ES cells to antero-lateral mesodermal cell, and that factor X promote differentiation from antero-lateral mesodermal cells to cardiomyocytes. Moreover, we found that Y-receptor is strongly expressed at the early differentiated cardiomyocytes, and administration of Y ligand promoted cell division in embryonic cardiomyocyte in vivo and early differentiated cardiomyocytes from ES cells. In combination with these factors, we can produce cardiomyocytes from marmoset and human ES cells. We also developed a novel method to purify cardiomyocytes from the mixture of cells by using mitochondria-specific incorporated dye. We transplanted the cardiomyocytes purified by this method, and the grafted cardiomyocytes can eliminate teratoma formation. Moreover, we also developed a novel method to enhance the cell survival after transplantation by a re-aggregation method. The grafted cell survival by needle injection into the heart was less than 1%, while this method can improve the efficiency up to 90%. We transplanted the purified cardiomyocytes into immuno-deficient mouse and common marmoset monkeys, In combination with these various techniques, we developed an efficient method to purify cardiomyocytes.

本研究试图建立一种从人类和普通狨猴 ES 细胞中产生、纯化和移植新心肌细胞的新方法。我们发现Noggin诱导ES细胞向前外侧中胚层细胞分化，而因子X促进前外侧中胚层细胞向心肌细胞分化。此外，我们发现Y受体在早期分化的心肌细胞中强烈表达，并且Y配体的施用促进了体内胚胎心肌细胞和来自ES细胞的早期分化心肌细胞的细胞分裂。结合这些因素，我们可以从狨猴和人类 ES 细胞中产生心肌细胞。我们还开发了一种新方法，通过使用线粒体特异性掺入染料从细胞混合物中纯化心肌细胞。我们移植了用这种方法纯化的心肌细胞，移植的心肌细胞可以消除畸胎瘤的形成。此外，我们还开发了一种通过重新聚集方法提高移植后细胞存活率的新方法。通过针头注射到心脏的移植细胞存活率不到1%，而这种方法可以将效率提高高达90%。我们将纯化的心肌细胞移植到免疫缺陷小鼠和普通狨猴体内，结合这些各种技术，我们开发了一种有效的纯化心肌细胞的方法。

项目成果

Transient and strong inhibition of BMP signals by Noggin induces cardiomyocyte differentiation in murine embryonic stem cells.

Noggin 对 BMP 信号的短暂强烈抑制可诱导小鼠胚胎干细胞中的心肌细胞分化。

• 发表时间: 2005
• 期刊: Nature Biotechnology 23
• 作者: Shnsuke Yuasa;Keiichi Fukuda;et al.
• 通讯作者: et al.

Chondromodulin-I mainstains cardiac valvular function by preventing angiogenesis.

软骨调节蛋白-I 通过阻止血管生成来维持心脏瓣膜功能。

• 发表时间: 2006
• 期刊: Nature Medicine 12
• 作者: Masatoyo Yoshioka;Keicihi Fukuda;et al.
• 通讯作者: et al.

Mesenchymal stem cell-derived cardiomyogenic cells. Cardiac Regeneration and Stem Cell Therapy.

间充质干细胞衍生的心肌细胞。

• 发表时间: 2006
• 期刊: edited by Leri A, Anversa P, Frishman WH. Blackwell Publishing. Oxford, UK.
• 作者: Keiichi Fukuda;Jun Fujita;Daihiko Hakuno;Shinji Makino.
• 通讯作者: Shinji Makino.

Wnt2 plays a key role in cardiac development via a non-canonical pathway.

Wnt2 通过非典型途径在心脏发育中发挥关键作用。

• 发表时间: 2008
• 作者: 0nizuka T;Yuasa S;Shimoji Ohno Y;Hattori F;Hara M;Hao C;Ogawa S;Fukuda K.
• 通讯作者: Fukuda K.

Cardiac neural crest cells contribute to the dormant multipotent stem cell in the mammalian heart.

• DOI: 10.1083/jcb.200504061
• 发表时间: 2005-09-26
• 期刊: The Journal of cell biology
• 作者: Tomita Y;Matsumura K;Wakamatsu Y;Matsuzaki Y;Shibuya I;Kawaguchi H;Ieda M;Kanakubo S;Shimazaki T;Ogawa S;Osumi N;Okano H;Fukuda K
• 通讯作者: Fukuda K