ヒトiPS細胞由来機能性臓器の試験管内誘導へ向けた集学的アプローチ

2014 Fiscal Year Annual Research Report

• Project Area: Hyper Bio Assembler for 3D Celluler Innovation
• Project/Area Number: 26106721
• Research Institution: Yokohama City University
• Principal Investigator: 武部 貴則 横浜市立大学, 医学部, 准教授 (20612625)
• Project Period (FY): 2014-04-01 – 2016-03-31
• Keywords: 再生医学

Outline of Annual Research Achievements

本研究では、申請者が独自開発したOrgan Bud Generation法を起点として、工学などさまざまな異分野の技術を融合し、発展型の培養系確立を目指している。
本年度では、同領域の5グループとの共同研究体制を構築した。A01班中内Gと連携のもと、造血細胞を新たに加えた器官原基形成法の検討を開始し、良好な結果を得つつある。さらに、A01班新井Gと共同で、器官原基の物理特性を評価し、その品質を定量する共同研究を開始した。一方、A02班 埼玉大学吉川Gと連携のもと、培養系における外部環境（生化学的/力学的誘導因子・培養基材の硬さ特性・培養期間など）の各種パラメータを検討し、その最適条件を解明することに成功した。成果は、権威ある学術誌であるCell Stem Cell誌に受理された。さらに、A03班鈴木Gと連携のもと、酸素透過膜を有したマイクロパターンプレートを用いることでヒトiPS細胞由来肝臓原基の効率的な分化誘導が可能であることを示した。また得られた肝臓原基を、A02班福田Gの高速血管モールディング系に導入することで、灌流培養を可能とする複合組織の創出に成功した。以上のように、A01班の新井Gとの連携など、当初計画を越えてさまざまな連携研究が生まれつつある。
今後も多角的な視点からブラッシュアップを継続していくことで、最終的にこの競争優位性の高いOrgan Bud Generation法を実用化技術へと発展させ、ヒトiPS細胞から高機能性の臓器を大量に、かつ、安価で、均一性と安全性を担保して創出可能な臓器構成方法を確立することを試みる。

Current Status of Research Progress

1: Research has progressed more than it was originally planned.

Reason

当初計画の実施内容は概ね完了した。特に、外部環境を至適化している埼玉大学との共同研究は、特許出願・論文発表など目覚ましい成果創出が達成された。さらに、名古屋大学新井Gと新たな共同研究を開始できたことなど、当初計画の想定を大幅に超える進捗状況であったといえる。

Strategy for Future Research Activity

iPS細胞の維持、肝前駆細胞分化誘導に必要な培地やサイトカイン等に必要な消耗品費に使用する。得られた細胞の評価のための、遺伝子発現解析試薬や免疫染色用抗体などの試薬にも経費が必要となる。また、研究の推進に必要な研究補助員の人件費、共同研究先への出張旅費などにも利用する。

Research Products

• [Journal Article] Transient vascularization of transplanted human adult-derived progenitors promotes self-organizing cartilage. (2014)
  • Author(s): Takebe T *, Kobayashi S, Suzuki H, Mizuno M, Chang YM, Yoshizawa E, Kimura M, Hori A, Asano J, Maegawa J, Taniguchi H
  • Journal Title: Journal of Clinical Investigation Volume: 124 Pages: 4325-4334
  • DOI: 10.1172/JCI76443
  • Peer Reviewed
• [Journal Article] Generation of a vascularized and functional human liver from an iPSC-derived organ bud transplant. (2014)
  • Author(s): Takebe T*, Zhang RR, Koike H, Kimura M, Yoshizawa E, Enomura M, Sekine K, Taniguchi H*
  • Journal Title: Nature Protocols Volume: 9 Pages: 396-409
  • DOI: 10.1038/nprot.2014.020
  • Peer Reviewed
• [Journal Article] Effect of cryopreservation and cell passage number on cell preparations destined for autologous chondrocyte transplantation. (2014)
  • Author(s): Nam BM, Kim BY, Jo YH, Lee S, Nemeno JG, Yang W, Lee KM, Kim H, Jang IJ, Takebe T, Lee JI
  • Journal Title: Transplant Proc Volume: 46 Pages: 1145-1149
  • DOI: 10.1016/j.transproceed.2013.11.117.
  • Peer Reviewed
• [Journal Article] Polycomb Group Protein Ezh2 Regulates Hepatic Progenitor Cell Proliferation and Differentiation in Murine Embryonic Liver. (2014)
  • Author(s): Koike H, Ouchi R, Ueno Y, Nakata S, Obana Y, Sekine K, Zheng YW, Takebe T, Isono K, Koseki H, Taniguchi H
  • Journal Title: PloS one Volume: 9 Pages: 0-1
  • DOI: 10.1371/journal.pone.0104776
  • Peer Reviewed
• [Journal Article] Effect of Preservation Conditions on Cartilage Tissue for Cell Transplantation (2014)
  • Author(s): Kim BY, Nam BM, Lee KM, Jo YH, Nemeno JG, Yang W, Lee S, Kim H, Jang IJ, Takebe T, Lee JI
  • Journal Title: Transplant Proc Volume: 46 Pages: 1139-1144
  • DOI: 10.1016/j.transproceed.2013.11.125.
  • Peer Reviewed
• [Journal Article] Artificial Islets From Hybrid Spheroids of Three Pancreatic Cell Lines. (2014)
  • Author(s): Jo YH, Jang IJ, Nemeno JG, Lee S, Kim BY, Nam BM, Yang W, Lee KM, Kim H, Takebe T, Kim YS, Lee JI
  • Journal Title: Transplant Proc Volume: 46 Pages: 1156-1160
  • DOI: 10.1016/j.transproceed.2013.11.074.
  • Peer Reviewed
• [Journal Article] Effects of Natural Cartilaginous Extracellular Matrix on Chondrogenic Potential for Cartilage Cell Transplantation (2014)
  • Author(s): Yang W, Lee S, Jo YH, Lee KM, Nemeno JG, Nam BM, Kim BY, Jang IJ, Kim HN, Takebe T, Lee JI
  • Journal Title: Transplant Proc Volume: 46 Pages: 1247-1250
  • DOI: 10.1016/j.transproceed.2013.11.082.
  • Peer Reviewed
• [Journal Article] Evidence of a Sophisticatedly Heterogeneous Population of Human Umbilical Vein Endothelial Cells (2014)
  • Author(s): Zheng YW, Nie YZ, Tsuchida T, Zhang, Aoki K, Sekine K, Ogawa M, Takebe T, Ueno Y, Sakakibara H, Hirahara F, Taniguchi H
  • Journal Title: Transplant Proc Volume: 46 Pages: 1251-1253
  • DOI: 10.1016/j.transproceed.2013.11.077.
  • Peer Reviewed
• [Journal Article] Fluorescent Labeling and Visualization of Human Induced Pluripotent Stem Cells With the Use of Transcription Activator–like Effector Nucleases. (2014)
  • Author(s): Sekine K, Takebe T, Taniguchi H
  • Journal Title: Transplant Proc Volume: 46 Pages: 1205-1207
  • Peer Reviewed
• [Journal Article] The CD133+CD44+ Precancerous Subpopulation of Oval Cells Is a Therapeutic Target for Hepatocellular Carcinoma. (2014)
  • Author(s): Zheng YW, Tsuchida T, Shimao T, Li B, Takebe T, Zhang RR, Sakurai Y, Ueno Y, Sekine K, Ishibashi N, Imajima M, Tanaka T, Taniguchi H
  • Journal Title: Stem Cells and Development Volume: 23 Pages: 2237-49
  • DOI: 10.1089/scd.2013.0577.
  • Peer Reviewed
• [Journal Article] Ring1B Promotes Hepatic Stem/Progenitor Cell Expansion via Simultaneous Suppression of Cdkn1a and Cdkn2a. (2014)
  • Author(s): Koike H, Ueno Y, Naito T, Shiina T, Ouchi R, Obana Y, Mori M, Sekine K, Takebe T, Zheng YW, Isono K, Koseki H, Taniguchi H
  • Journal Title: Hepatology Volume: 60 Pages: 323-333
  • DOI: 10.1002/hep.27046.
  • Peer Reviewed
• [Journal Article] The development of humanized liver with Rag1 knockout rats. (2014)
  • Author(s): Tsuchida T, Zheng YW, Zhang RR, Takebe T, Ueno Y, Sekine K, Taniguchi H
  • Journal Title: Transplant Proc Volume: 46 Pages: 1191-1193
  • DOI: 10.1016/j.transproceed.2013.12.026.
  • Peer Reviewed
• [Journal Article] Tropism of cancer stem cells to a specific distant organ. (2014)
  • Author(s): Okuda R, Sekine K, Hisamatsu D, Ueno Y, Takebe T, Zheng YW, Taniguchi H
  • Journal Title: In Vivo Volume: 28 Pages: 361-365
  • Peer Reviewed
• [Journal Article] Elastic cartilage reconstruction by transplantation of cultured hyaline cartilage-derived chondrocytes. (2014)
  • Author(s): Mizuno M, Takebe T*, Kobayashi S, Kimura S, Masutani M, Lee S, Jo YH, Lee JI, Taniguchi H*
  • Journal Title: Transplant Proc Volume: 46 Pages: 1217-1221
  • DOI: 10.1016/j.transproceed.2013.12.006.
  • Peer Reviewed
• [Journal Article] Identification of Proliferating Human Hepatic Cells from Human Induced Pluripotent Stem Cells. (2014)
  • Author(s): Zhang RR Takebe T*, Sekine K, Koike H, Zheng YW Taniguchi H*
  • Journal Title: Transplant Proc Volume: 46 Pages: 1201-1204
  • DOI: 10.1016/j.transproceed.2013.12.021.
  • Peer Reviewed
• [Journal Article] Engineering of human hepatic tissue with functional vascular networks. (2014)
  • Author(s): Takebe T*, Koike N*, Sekine K, Fujiwara R, Amiya T, Zheng YW, Taniguchi H*
  • Journal Title: Organogenesis Volume: 10 Pages: 260-267
  • DOI: 10.4161/org.27590
  • Peer Reviewed
• [Journal Article] High-resolution intravital imaging for monitoring the transplanted islet in mice. (2014)
  • Author(s): Takahashi Y, Takebe T*, Enomura M, Koike N, Lee S, Nemeno JG, Sekine K, Lee JI, Taniguchi H*
  • Journal Title: Transplant Proc Volume: 46 Pages: 1166-1168
  • DOI: 10.1016/j.transproceed.2013.11.089.
  • Peer Reviewed
• [Journal Article] Self-Formation of Vascularized Hepatic Tissue from Human Adult Hepatocyte. (2014)
  • Author(s): Enomoto Y, Enomura M, Takebe T*, Mitsuhashi Y, Kimura M, Yoshizawa E, Taniguchi H*
  • Journal Title: Transplant Proc Volume: 46 Pages: 1243-1246
  • DOI: 10.1016/j.transproceed.2013.11.086.
  • Peer Reviewed
• [Presentation] Liver organogenesis (2015)
  • Author(s): Takebe T
  • Organizer: ASTELLAS FRANCE Annual Meeting of Transplantation Club
  • Place of Presentation: Lille, France
  • Year and Date: 2015-02-06 – 2015-02-06
  • Invited
• [Presentation] Transient vascularization of transplanted human adult-derived progenitors promotes self-organizing cartilage. (2014)
  • Author(s): Takebe T
  • Organizer: Asian Cartilage Repair Society 2nd Annual Congress
  • Place of Presentation: Seoul, Korea
  • Year and Date: 2014-12-07 – 2014-12-07
  • Invited
• [Presentation] 臨床応用へ向けたiPS細胞由来ヒト臓器原基操作技術の開発 (2014)
  • Author(s): 武部貴則
  • Organizer: 第41回日本臓器保存生物医学会学術集会
  • Place of Presentation: 千里ライフサイエンスセンター（大阪府）
  • Year and Date: 2014-11-28 – 2014-11-28
  • Invited
• [Presentation] 器官発生初期プロセスの再現による複雑なヒト臓器の人為的構成 (2014)
  • Author(s): 武部貴則
  • Organizer: 第37回日本分子生物学会年会シンポジウム
  • Place of Presentation: パシフィコ横浜（神奈川県）
  • Year and Date: 2014-11-25 – 2014-11-27
  • Invited
• [Presentation] Realization of iPSC-Organ Bud Transplantation Therapy. (2014)
  • Author(s): Takebe T
  • Organizer: World Alliance forum
  • Place of Presentation: San Francisco, CA, USA
  • Year and Date: 2014-11-06 – 2014-11-07
  • Invited
• [Presentation] Vascularized and functional human liver from an iPSC-derived organ bud transplant. (2014)
  • Author(s): Takebe T
  • Organizer: Advances and Applications of Functional Hepatocytes Symposium
  • Place of Presentation: Shanghai, China.
  • Year and Date: 2014-10-29 – 2014-10-30
  • Invited
• [Presentation] Pluripotent stem cells and organ reconstitution for rare diseases. (2014)
  • Author(s): Takebe T
  • Organizer: Translational Science of Rare Diseases- From Rare to Care II
  • Place of Presentation: Munich, Germany
  • Year and Date: 2014-10-08 – 2014-10-10
  • Invited
• [Presentation] 多細胞系からなる複雑なヒト臓器の人為的構成 (2014)
  • Author(s): 武部貴則
  • Organizer: 第66回日本生物工学会大会
  • Place of Presentation: 札幌コンベンションセンター（北海道）
  • Year and Date: 2014-09-10 – 2014-09-10
  • Invited
• [Presentation] Vascularized and functional human liver from an iPSC-derived organ bud transplant. (2014)
  • Author(s): Takebe T
  • Organizer: KEY Forum: From Stem Cells to Organs
  • Place of Presentation: 熊本市医師会館（熊本県）
  • Year and Date: 2014-09-04 – 2014-09-05
  • Invited
• [Presentation] Vascularized and functional human liver from an iPSC-derived organ bud transplant. (2014)
  • Author(s): Takebe T
  • Organizer: 47th Annual Meeting for the Japanese Society of Developmental Biologists
  • Place of Presentation: WINC AICHI（愛知県）
  • Year and Date: 2014-05-28 – 2014-05-28
  • Invited
• [Presentation] Realization of human iPSC-derived organ bud transplantation therapy. (2014)
  • Author(s): Takebe T
  • Organizer: The Whole Liver Replacement State-of-the-Science Summit
  • Place of Presentation: Chicago, USA
  • Year and Date: 2014-04-29 – 2014-04-30
  • Invited
• [Presentation] Vascularized and functional human liver from an iPSC-derived organ bud transplant. (2014)
  • Author(s): Takebe T
  • Organizer: 7th Pan Pacific Symposium on Stem Cells and Cancer Research (PPSSC)
  • Place of Presentation: Taichung, Taiwan
  • Year and Date: 2014-04-12 – 2014-04-14
  • Invited
• [Book] iPS細胞由来器官原基の人為的構成に基づくヒト肝臓の創出 (2014)
  • Author(s): 武部貴則、谷口英樹
  • Total Pages: 4
  • Publisher: 横浜医学
• [Book] 幹細胞と微小環境の相互作用に基づくヒト3次元組織の人為的再構成 (2014)
  • Author(s): 武部貴則、谷口英樹
  • Total Pages: 5
  • Publisher: がん分子標的治療
• [Book] 多能性幹細胞を用いた膵β細胞分化誘導研究の最新動向 (2014)
  • Author(s): 高橋禎暢、武部貴則、谷口英樹
  • Total Pages: 8
  • Publisher: Organ Biology
• [Book] Novel strategies for liver therapy using stem cells. (2014)
  • Author(s): Rashid T, Takebe T, Nakauchi H
  • Total Pages: 3
  • Publisher: Gut
• [Book] Human iPSC-derived miniature organs: a tool for drug studies. (2014)
  • Author(s): Takebe T, Taniguchi H
  • Total Pages: 3
  • Publisher: Clinical Pharmacology Therapeutics
• [Book] ヒトiPS細胞を活用した再生医療の開発戦略 (2014)
  • Author(s): 谷口英樹、武部貴則
  • Total Pages: 2
  • Publisher: 消化器外科
• [Book] ヒト代謝性臓器の創出に向けた開発戦略 (2014)
  • Author(s): 谷口英樹、武部貴則
  • Total Pages: 9
  • Publisher: 最新医学
• [Book] ヒト臓器の人為的構成に基づく肝細胞の分化誘導 ES・iPS細胞実験スタンダード―再生・創薬・疾患研究のプロトコールと臨床応用の必須知識― (2014)
  • Author(s): 14.武部貴則、関根圭輔、谷口英樹
  • Total Pages: 10
  • Publisher: 羊土社
• [Book] Human iPSC-Derived Miniature Organs: A Tool for Drug Studies. (2014)
  • Author(s): Takebe T*, Taniguchi H
  • Total Pages: 3
  • Publisher: Clinical Pharmacology & Therapeutics
• [Patent(Industrial Property Rights)] 原始腸内胚葉細胞及びその作製方法 (2014)
  • Inventor(s): 武部貴則(40%)、張冉冉(10%)、谷口英樹(50%)
  • Industrial Property Rights Holder: 武部貴則(40%)、張冉冉(10%)、谷口英樹(50%)
  • Industrial Property Rights Type: 特許
  • Industrial Property Number: 2014-248694
  • Filing Date: 2014-12-08
• [Patent(Industrial Property Rights)] 培養方法及び細胞塊 (2014)
  • Inventor(s): 江尻洋子(10%)、谷口英樹(50%)、 武部貴則(40%)
  • Industrial Property Rights Holder: 江尻洋子(10%)、谷口英樹(50%)、 武部貴則(40%)
  • Industrial Property Rights Type: 特許
  • Industrial Property Number: 2014-112959
  • Filing Date: 2014-05-29