Establishment of human pluripotent stem cell-derived pace-making cells using physiological approach and its application to the regenerative medicine

生理学方法建立人多能干细胞来源的起搏细胞及其在再生医学中的应用

• 批准号: 23659112
• 负责人: HISATOME Ichiro
• 金额: $ 2.41万
• 依托单位: Tottori University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Challenging Exploratory Research
• 财政年份: 2011
• 资助国家: 日本
• 起止时间: 2011 至 2012
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-23659112/
• 关键词: human ES cells; HCN4; pacemaking cells; ヒトES細胞; ヒトiPS細胞; HCN4搭載BACベクター; ヒトＥＳ細胞; ペースメーカ細胞

We attempted to establish the human ES cells harboringHCN4-GFP BAC vector to isolate human pluripotent stem cell-derived pace-making cells using physiological approach. We construct BAC vector harboring the knocked GFP gene into the promoter region of HCN4 gene and introduced it into human ES cells (KhES-1) to establish the several clones including clone #1. These clones expressed the pluripotent gene markers and normal karyo type of chromosome. The embryoid body derived from clone#1 to differentiate cardiac cells included the HCN4-GFP positive cells at 5% out of total human ES cells. HCN4-GFP positive cells expressed cardiac contractile proteins and HCN4 channels. HCN4-GFP positive cells expressed the several ion channels being responsible for their automaticity. The Ca transient derived from the aggregation of HCN4-GFP positive cells propagated into the HL-1 atrial cell-sheet. Taken together, we establish the human ES cells harboringHCN4-GFP BAC vector and successfully isolate the HCN4-GFP positive pacemakingcells.

我们试图利用生理学方法建立携带hcn4 - gfp BAC载体的人胚胎干细胞，以分离人多能干细胞衍生的起搏细胞。我们在HCN4基因启动子区构建含有敲除GFP基因的BAC载体，并将其导入人ES细胞（KhES-1），建立了包括克隆1在内的多个克隆。这些克隆表达了多能性基因标记和正常核型染色体。从克隆1中获得的胚胎样体分化心脏细胞包括HCN4-GFP阳性细胞，占人类胚胎干细胞总数的5%。HCN4- gfp阳性细胞表达心脏收缩蛋白和HCN4通道。HCN4-GFP阳性细胞表达了负责其自动性的几个离子通道。由HCN4-GFP阳性细胞聚集产生的Ca瞬态传播到HL-1心房细胞片。综上所述，我们建立了携带HCN4-GFP BAC载体的人ES细胞，并成功分离出HCN4-GFP阳性起搏细胞。

项目成果

Functional integration of HCN4 positive cardiac pacemaking cells derived from mouse embryonicstem cells with recipient cardiomyocytes

小鼠胚胎干细胞来源的 HCN4 阳性心脏起搏细胞与受体心肌细胞的功能整合

• 发表时间: 2013
• 作者: 宮川桃子;石龍徳;内山安男;太田啓介;Kumi Morikawa
• 通讯作者: Kumi Morikawa

Enhancing effects of salicylate on quinidine-induced block of human wild type and LQT3 related mutant cardiac Na+ channels.

水杨酸盐对奎尼丁诱导的人类野生型和 LQT3 相关突变心脏 Na 通道阻断的增强作用。

• DOI: 10.2220/biomedres.32.303
• 发表时间: 2011
• 期刊: Biomedical research
• 作者: T. Urashima;Y. Kurata;J. Miake;Masaru Kato;Kazuyoshi Ogura;A. Yano;M. Adachi;Yasunori Tanaka;K. Yamada;T. Hamada;E. Mizuta;M. Kuwabara;Masahiko Kato;Yasutaka Yamamoto;K. Ogino;Akio Yoshida;Y. Shirayoshi;I. Hisatome
• 通讯作者: I. Hisatome

Transcriptional activation of SAP97 by HSF-1 stabilizes Kv1. 5 in HL-1 cells.

HSF-1 对 SAP97 的转录激活可稳定 Kv1。

• 发表时间: 2011
• 期刊: Brit. J. Pharmacol
• 作者: Y. K. Ting;A. Nakai(他15名、12番目)
• 通讯作者: A. Nakai(他15名、12番目)

Udin Bahrudin Carbohydrate diet links to higher risk of significant coronary artery disease in young Indonesian patients Cardiometabolic Investigation study

Udin Bahrudin 碳水化合物饮食与印度尼西亚年轻患者患严重冠状动脉疾病的较高风险有关

• 发表时间: 2012
• 期刊: Biomedical Research
• 作者: Sugiri;Sefri Noventi;Ichiro Hisatome
• 通讯作者: Ichiro Hisatome

Intracelluar Ca2+ handling regulates the automaticity of HCN4-GFP(+) pacemaking cells derived frommurine ES cells during cardiac differentiation

细胞内 Ca2 处理调节源自小鼠 ES 细胞的 HCN4-GFP( ) 起搏细胞在心脏分化过程中的自动性

• 发表时间: 2013
• 作者: 藤原祐一郎;マルタペレス;黒川竜紀;ピータラルソン;岡村康司;Udin Bahrudin
• 通讯作者: Udin Bahrudin