Isolation and characterization of cardiac pacemaking cells derived from human embryonic stem cells

• Project/Area Number: 25893134
• Research Category: Grant-in-Aid for Research Activity Start-up
• Allocation Type: Single-year Grants
• Research Field: General physiology
• Research Institution: Tottori University
• Principal Investigator: MORIKAWA Kumi 鳥取大学, 医学部附属病院, 助教 (90707217)
• Project Period (FY): 2013-08-30 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥2,730,000 (Direct Cost: ¥2,100,000、Indirect Cost: ¥630,000); Fiscal Year 2014: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2013: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
• Keywords: ペースメーカ細胞 / ヒトES細胞 / HCN4 / Ifチャネル / 分化誘導 / 心筋細胞 / 自動能 / 可視化 / ペースメーカ / イオンチャネル / 心臓

Outline of Final Research Achievements

The difficulty of preparation in vivo hampered us to analyze directly human sino-atrial pacemaker cells. Therefore we have established the experimental system for preparation of human pacemaker cells derived from human embryonic stem cells (hESCs). In this system, HCN4 positive cells were specifically visualized with CFP (cyan fluorescent protein). The sorted CFP positive cells from hESCs expressed HCN4 maker gene and displayed If current, which could resulted in the automaticity of those cells. Thus CFP positive cells from differentiating hESCs have the essentially same properties with sino-atrial pacemaker cells, including the gene expression profiles, electrophysiological properties. In addition, we targeted mCherry to the MLC2v locus, a good maker for ventricular cells, in the HCN4CFP-BAC hESCs by using CRIPER/CAS9 system. Currently we purified and analyzed HCN4 positive pacemaker cells and MLC2v positive ventricular cells to characterize the cardiac cells with human origin.

Research Products

• [Journal Article] <b>Adipose stem cell sheets improved cardiac function in the rat myocardial infarction, but did not alter cardiac contractile responses to β-adrenergic stim</b><b>ulation </b> (2015)
  • Author(s): Otsuki Y, Nakamura Y, Harada S, Yamamoto Y, Ogino K, Morikawa K, Ninomiya H, Miyagawa S, Sawa Y, Hisatome I, Nishimura M.
  • Journal Title: Biomedical Research Volume: 36 Issue: 1 Pages: 11-19
  • DOI: 10.2220/biomedres.36.11
  • NAID: 130004903877
  • ISSN: 0388-6107, 1880-313X
  • Peer Reviewed / Open Access
• [Journal Article] <b>VEGF secretion by adipose tissue-derived regenerative cells is impaired under hyperglycemic conditions via glucose transporter activation and ROS incr</b><b>ease </b> (2014)
  • Author(s): Matsugami H, Harada Y, Kurata Y, Yamamoto Y, Otsuki Y, Yaura H, Inoue Y, Morikawa K, Yoshida A, Shirayoshi Y, Suyama Y, Nakayama B, Iwaguro H, Yamamoto K, Hisatome I.
  • Journal Title: Biomedical Research Volume: 35 Issue: 6 Pages: 397-405
  • DOI: 10.2220/biomedres.35.397
  • NAID: 130004903869
  • ISSN: 0388-6107, 1880-313X
  • Peer Reviewed / Open Access
• [Journal Article] Instability of KCNE1-D85N that Causes Long QT Syndrome: Stabilization by Verapamil (2014)
  • Author(s): Sakata S, Kurata Y, Li P, Notsu T, Morikawa K, Miake J, Higaki K, Yamamoto Y, Yoshida A, Shirayoshi Y, Yamamoto K, Horie M, Ninomiya H, Kanzaki S, Hisatome I.
  • Journal Title: Pacing Clinical Electrophysiology Volume: Epub ahead of print Issue: 7 Pages: 1-11
  • DOI: 10.1111/pace.12360
  • Peer Reviewed / Open Access
• [Journal Article] Bepridil Suppresses Apoptosis in HL-1 Cardiac Atrial Myocytes Expressing Mutant E334K cMyBPC. (2013)
  • Author(s): Endo R, Notsu T, Mishima M, Morikawa K, Li P, Ikeda N, Ninomiya H, Shirayoshi Y, Hisatome I.
  • Journal Title: Yonago Acta Medica Volume: 56(4) Pages: 93-95
  • NAID: 120005350605
  • Peer Reviewed
• [Journal Article] Hsp90 prevents interaction between CHIP and HERG proteins to facilitate maturation of wild-type and mutant HERG proteins (2013)
  • Author(s): Iwai C, P Li,
  • Journal Title: Cardiovasc Res Volume: ３ Issue: 3 Pages: 520-528
  • DOI: 10.1093/cvr/cvt200
  • Peer Reviewed
• [Journal Article] Simultaneous Treatment with Azelnidipine and Olmesartan Inhibits Apoptosis of Hl-1 Cardiac Myocytes Expressing E334k cMyBPC (2013)
  • Author(s): Bahrudin U, Ikeda N, Utami SB, Maharani N, Morikawa K, Li P
  • Journal Title: Drug Res (Stuttg). Volume: １０ Issue: 10 Pages: 515-520
  • DOI: 10.1055/s-0033-1347188
  • Peer Reviewed
• [Journal Article] 胚性幹細胞からの機能的な甲状腺の作出 (2013)
  • Author(s): 森川久未, 久留一郎, 吉田明雄
  • Journal Title: 日本甲状腺学会雑誌 Volume: 4(1) Pages: 68-69
• [Journal Article] 脂肪由来幹細胞による再生医療 (2013)
  • Author(s): 森川久未, 久留一郎
  • Journal Title: 鳥取県西部医師会報 Volume: 170 Pages: 23-25
• [Journal Article] AMP deaminase 3 plays a critical role in remote reperfusion lung injury (2013)
  • Author(s): Li P, Ogino K, Morikawa K, Shirayoshi Y, Hisatome I, et al.
  • Journal Title: Biochem Biophys Res Commun. Volume: 431(2) Issue: 1 Pages: 131-6
  • DOI: 10.1016/j.bbrc.2013.03.056
  • Peer Reviewed
• [Presentation] ISOLATION AND CHARACTERIZATION OF HCN4 POSITIVE CELLS DERIVED FROM HUMAN EMBRYONIC STEM CELLS (2015)
  • Author(s): Morikawa K, Nozaki D, Shirayoshi Y, Hisatome I.
  • Organizer: International Society for Stem Cell Research 13th Annual Meeting
  • Place of Presentation: Stockholm, Sweden
  • Year and Date: 2015-06-24 – 2015-06-27
• [Presentation] 生物学的ペースメーカ細胞による再生医療を目指して (2015)
  • Author(s): 森川 久未
  • Organizer: 第7回再生医療学フォーラム in 山陰
  • Place of Presentation: 米子全日空ホテル, 鳥取
  • Year and Date: 2015-05-25
  • Invited
• [Presentation] HCN4 Positive Cells Derived from Pluripotent Stem Cells Show Automaticity and Pacemaking Ability (2015)
  • Author(s): Shirayoshi Y, Morikawa K, Miake J, Hisatome I
  • Organizer: The 79th Annual Scientific Meeting of the Japanese Circulation Society
  • Place of Presentation: 大阪国際会議場, 大阪
  • Year and Date: 2015-04-24 – 2015-04-26
• [Presentation] 高血糖は糖輸送体を介して脂肪由来間葉系幹細胞の血管新生能を障害する (2015)
  • Author(s): 松上 紘生, 原田 雄輔, 森川 久未, 山本 康孝, 陶山 淑子, 中山 敏, 吉田 明雄, 白吉 安昭, 齋藤 憲輝, 久留 一郎
  • Organizer: 第14回日本再生医療学会総会
  • Place of Presentation: パシフィコ横浜, 神奈川
  • Year and Date: 2015-03-19 – 2015-03-21
• [Presentation] 乳房再建治療メカニズムの解明に向けた乳房再建モデルマウス作製の試み (2015)
  • Author(s): 森川 久未, 八浦 妃佐子, 陶山 淑子, 中山 敏, 久留 一郎
  • Organizer: 第14回日本再生医療学会総会
  • Place of Presentation: パシフィコ横浜, 神奈川
  • Year and Date: 2015-03-19 – 2015-03-21
• [Presentation] Electrical Properties of Human iPS Cells-derived Cardiac Myocytes from the Patient with Long QT Syndrome Harboring a Novel Mutation of KCNQ1 (2014)
  • Author(s): Sogo T, Ichinose T, Morikawa K, Ikeda N, Shirayoshi Y, Kurata Y, Miake J, Yamamoto K, Yuasa S, Fukuda K, Hisatome I
  • Organizer: 第29回 日本不整脈学会・第31回 日本心電学会 合同学術大会
  • Place of Presentation: 東京プリンスホテル, 東京
  • Year and Date: 2014-07-22 – 2014-07-25
• [Remarks] 鳥取大学大学院 医学系研究科 機能再生医科学専攻 遺伝子再生医療学講座 再生医療学部門
  • URL: http://www.med.tottori-u.ac.jp/regmed/
• [Remarks] 鳥取大学医学部附属病院 次世代高度医療推進センター
  • URL: http://www2.hosp.med.tottori-u.ac.jp/departments/center/next-generation-advanced-medical-promotion-center/
• [Remarks] 生物学的心臓ペースメーカー細胞の作成とその応用
  • URL: http://www.med.tottori-u.ac.jp/introduction/grad/297/430/563/1345.html
• [Remarks] 鳥取大学大学院医学系研究科 機能再生医科学専攻 遺伝子再生医療学講座再生医療学部門
  • URL: http://www.med.tottori-u.ac.jp/regmed/
• [Remarks] 次世代高度医療推進センター｜鳥取大学医学部附属病院
  • URL: http://www2.hosp.med.tottori-u.ac.jp/departments/center/next-generation-advanced-medical-promotion-center/