薬物代謝のリアルタイム評価を可能とするマイクロ構造を備えた肝組織の作製

• Project Area: Hyper Bio Assembler for 3D Celluler Innovation
• Project/Area Number: 26106722
• Research Category: Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area)
• Allocation Type: Single-year Grants
• Review Section: Science and Engineering
• Research Institution: Yokohama City University
• Principal Investigator: 小島 伸彦 横浜市立大学, 総合科学部, 准教授 (90342956)
• Project Period (FY): 2014-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥9,360,000 (Direct Cost: ¥7,200,000、Indirect Cost: ¥2,160,000); Fiscal Year 2015: ¥2,470,000 (Direct Cost: ¥1,900,000、Indirect Cost: ¥570,000); Fiscal Year 2014: ¥6,890,000 (Direct Cost: ¥5,300,000、Indirect Cost: ¥1,590,000)
• Keywords: 肝臓 / 類洞 / 肝細胞索 / 毛細胆管 / スフェロイド / 三次元培養 / 組織工学 / 再生医療

Outline of Annual Research Achievements

最終年度である本年度は、昨年度より計画の達成がずれ込んでいた計画１：「類洞・肝細胞索に加えて、毛細胆管構造を備えた『微細構造化３次元肝組織』の構築」を実施し、さらに計画２：『「薬物の吸収・代謝・排泄をリアルタイムに評価できるデバイス」の作製』と計画３：『薬物代謝に関する遺伝子ノックダウンによる「薬物代謝デバイスの有効性」の検証』に取り組んだ。
計画１について、マウス胎仔肝細胞とハイドロゲルビーズを使って類洞構造および肝細胞索構造を持つ肝スフェロイドを作製し、その後比較的長期間の培養を行うことによって毛細胆管と想定される構造の構築に成功した。計画２について、モデル薬物の吸収・代謝・排泄を共焦点顕微鏡下で観察できるデバイスの作製を行うことができた。計画３について、RNAiによるノックダウンよりも簡便で確実な手法としてトランスポーター阻害剤を用いた評価法を行い、毛細胆管と思われる構造への排泄能力の低下を観測できた。
また、超高速バイオアセンブラの取り組みとして、膵島や骨髄、さらには心筋や精巣の再構築にも取り組み、成果を得た。

Research Progress Status

27年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

27年度が最終年度であるため、記入しない。

Research Products

• [Journal Article] Fabrication of bone marrow-like tissue in vitro from dispersed-state bone marrow cells. (2016)
  • Author(s): Sayo, K., Aoki, S. and Kojima, N.
  • Journal Title: Regenerative Therapy Volume: 3 Pages: 32-37
  • DOI: 10.1016/j.reth.2016.01.008
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] β-galactoside-mediated tissue organization during islet reconstitution (2016)
  • Author(s): Kamitori, S., Ozeki, Y. and Kojima, N.
  • Journal Title: Regenerative Therapy Volume: 3 Pages: 11-14
  • DOI: 10.1016/j.reth.2016.01.006
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Induction of hepatic tissues in multicellular spheroids composed of murine fetal hepatic cells and embedded hydrogel beads. (2016)
  • Author(s): Motoyama, W., Sayo, K., Mihara, H., Aoki, S. and Kojima, N.
  • Journal Title: Regenerative Therapy Volume: 3 Pages: 7-10
  • DOI: 10.1016/j.reth.2016.01.007
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] The mechanism and control of Jagged1 expression in Sertoli cells. (2016)
  • Author(s): Okada, R., Hara, T., Sato, T., Kojima, N. and Nishina, Y.
  • Journal Title: Regenerative Therapy Volume: 3 Pages: 75-81
  • DOI: 10.1016/j.reth.2016.02.005
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Non-neural and cardiac differentiating properties of Tbx6-expressing mouse embryonic stem cells. (2016)
  • Author(s): Yano, Y., Iimura, N., Kojima, N. and Uchiyama, H.
  • Journal Title: Regenerative Therapy Volume: 3 Pages: 1-6
  • DOI: 10.1016/j.reth.2016.02.001
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Presentation] 高付加価値なスフェロイドをつくる技術 (2016)
  • Author(s): 小島伸彦
  • Organizer: 日本動物実験代替法学会 第29回大会
  • Place of Presentation: 九州大学百年講堂＆同窓会館
  • Year and Date: 2016-11-16
  • Invited
• [Presentation] A method to fabricate multicellular spheroids containing various amount of extracellular matrix (2016)
  • Author(s): Tao, F. and Kojima, N.
  • Organizer: TERMIS-EU 2016
  • Place of Presentation: Uppsala Concert and Congress, Uppsala, Sweden
  • Year and Date: 2016-06-28
  • Int'l Joint Research
• [Presentation] Effect of SCF and TPO on the bone marrow-like tissue (2016)
  • Author(s): Sayo, K., Aoki, S. and Kojima, N.
  • Organizer: TERMIS-EU 2016
  • Place of Presentation: Uppsala Concert and Congress, Uppsala, Sweden
  • Year and Date: 2016-06-28
  • Int'l Joint Research
• [Presentation] Inhibition of epithelial-mesenchymal transition in cholangiocyte 3D culture by embedding alginate hydrogel beads (2016)
  • Author(s): Asakura, Y. and Kojima, N.
  • Organizer: TERMIS-EU 2016
  • Place of Presentation: Uppsala Concert and Congress, Uppsala, Sweden
  • Year and Date: 2016-06-28
  • Int'l Joint Research
• [Presentation] 肝スフェロイド内部の微細構造を制御する技術の開発 (2016)
  • Author(s): 小島伸彦
  • Organizer: 第55回日本生体医工学会大会
  • Place of Presentation: 富山国際会議場
  • Year and Date: 2016-04-28
• [Presentation] 疾患モデル作製に向けた骨髄様組織の再構築 (2016)
  • Author(s): 佐用かなえ，青木茂久，小島伸彦
  • Organizer: 細胞アッセイ研究会
  • Place of Presentation: 東京大学生産技術研究所コンベンションホール
  • Year and Date: 2016-01-19
• [Presentation] ECM含有スフェロイド作製のための新規技術 (2016)
  • Author(s): 田尾文哉，小島伸彦
  • Organizer: 細胞アッセイ研究会
  • Place of Presentation: 東京大学生産技術研究所コンベンションホール
  • Year and Date: 2016-01-19
• [Presentation] 細胞外マトリクスの充填による三次元細胞凝集体の機能改変（A method to form ECM-loaded spheroids） (2015)
  • Author(s): 田尾文哉，小島伸彦
  • Organizer: 第38回日本分子生物学会年会
  • Place of Presentation: 神戸ポートアイランド
  • Year and Date: 2015-12-02
• [Presentation] 分散状態の骨髄細胞を用いた３次元的な骨髄様組織の作製（Reconstruction of the 3D BM-like tissue from dispersed-state BM cells） (2015)
  • Author(s): 佐用かなえ，青木茂久，小島伸彦
  • Organizer: 第38回日本分子生物学会年会
  • Place of Presentation: 神戸ポートアイランド
  • Year and Date: 2015-12-02
• [Presentation] A New Method for Fabricating Cell-embedded ECM Capsules and ECM-loaded Spheroids. (2015)
  • Author(s): Tao, F., Aoki, S. and Kojima, N.
  • Organizer: 26th IEEE International Symposium on Micro-Nano Mechatronics and Human Science in 2015 (MHS2015)
  • Place of Presentation: Noyori Memorial Conference Hall, Nagoya University, Aichi, Japan
  • Year and Date: 2015-11-24
  • Int'l Joint Research
• [Presentation] A Method to Reorganize the Bone Marrow-like Tissue with Suspension of Bone Marrow cells. (2015)
  • Author(s): Kanae, S., Aoki, S. and Kojima, N.
  • Organizer: 26th IEEE International Symposium on Micro-Nano Mechatronics and Human Science in 2015 (MHS2015)
  • Place of Presentation: Noyori Memorial Conference Hall, Nagoya University, Aichi, Japan
  • Year and Date: 2015-11-24
  • Int'l Joint Research
• [Presentation] Rapid Formation of the Bone Marrow-like Tissue (2015)
  • Author(s): Sayo, K., Aoki, S. and Kojima, N.
  • Organizer: International Conference on Biofabrication 2015
  • Place of Presentation: Utrecht University Hall, Utrecht, Netherlands
  • Year and Date: 2015-11-08
  • Int'l Joint Research
• [Presentation] Prevention of apoptosis in epithelial-cell-spheroids (2015)
  • Author(s): Motoyama, F., Aoki, S. and Kojima, N.
  • Organizer: International Conference on Biofabrication 2015
  • Place of Presentation: Utrecht University Hall, Utrecht, Netherlands
  • Year and Date: 2015-11-08
  • Int'l Joint Research
• [Presentation] A new method for fabricating cell-embedded ECM capsules and ECM-loaded spheroids (2015)
  • Author(s): Tao, F. and Kojima, N.
  • Organizer: International Conference on Biofabrication 2015
  • Place of Presentation: Utrecht University Hall, Utrecht, Netherlands
  • Year and Date: 2015-11-08
  • Int'l Joint Research
• [Presentation] 細胞社会の特性を利用した組織微細構造の再構築 (2015)
  • Author(s): 小島伸彦
  • Organizer: 第19回日本内分泌病理学会学術総会
  • Place of Presentation: 佐賀県・アバンセ
  • Year and Date: 2015-10-25
  • Invited
• [Presentation] 3D Microfabrication and Cell Polarity Formation in Multicellular Spheroids (2015)
  • Author(s): Motoyama, Y., Aoki, S. and Kojima, N.
  • Organizer: 4th TERMIS World Congress 2015
  • Place of Presentation: Boston Marriott Copley Place Hotel, Boston, Massachusetts, USA.
  • Year and Date: 2015-09-09
  • Int'l Joint Research
• [Presentation] The Bone Marrow-like Tissue In Vitro: Rapid Aggregation and Reconstruction of Dispersed Bone Marrow Cells (2015)
  • Author(s): Sayo, K., Aoki, S. and Kojima, N.
  • Organizer: 4th TERMIS World Congress 2015
  • Place of Presentation: Boston Marriott Copley Place Hotel, Boston, Massachusetts, USA.
  • Year and Date: 2015-09-09
  • Int'l Joint Research
• [Presentation] Role of sugar chain in the self-organization of the pancreatic islet structure in vitro (2015)
  • Author(s): Kamitori, S., Ozeki, Y. and Kojima, N.
  • Organizer: 4th TERMIS World Congress 2015
  • Place of Presentation: Boston Marriott Copley Place Hotel, Boston, Massachusetts, USA.
  • Year and Date: 2015-09-08
  • Int'l Joint Research
• [Presentation] Enhancement of Albumin Secretion of Hepatocytes by Cholangiocytes and Hydrogel Beads in 3D culture (2015)
  • Author(s): Asakura, Y. and Kojima, N.
  • Organizer: 2015 Taiwan-Japan Forum on Additive Manufacturing and Bioengineering
  • Place of Presentation: National Formosa University, Yunlin, Taiwan
  • Year and Date: 2015-09-01
  • Int'l Joint Research
• [Presentation] Prevention of Apoptosis in Epithelial Cell Spheroids (2015)
  • Author(s): Motoyama, W., Aoki, S. and Kojima, N.
  • Organizer: 2015 Taiwan-Japan Forum on Additive Manufacturing and Bioengineering
  • Place of Presentation: National Formosa University, Yunlin, Taiwan
  • Year and Date: 2015-09-01
  • Int'l Joint Research
• [Presentation] Control of Islet Organization in Vitro (2015)
  • Author(s): Kamitori, S., Ozeki, Y. and Kojima, N.
  • Organizer: 2015 Taiwan-Japan Forum on Additive Manufacturing and Bioengineering
  • Place of Presentation: National Formosa University, Yunlin, Taiwan
  • Year and Date: 2015-09-01
  • Int'l Joint Research
• [Presentation] Cell-hydrogel Complexes Showing Liver Function (2015)
  • Author(s): Tao, F. and Kojima, N.
  • Organizer: 2015 Taiwan-Japan Forum on Additive Manufacturing and Bioengineering
  • Place of Presentation: National Formosa University, Yunlin, Taiwan
  • Year and Date: 2015-09-01
  • Int'l Joint Research
• [Presentation] In Vitro Reconstruction of the Seminiferous Tubles (2015)
  • Author(s): Yamada, A., Okada, R., Nishina, Y. and Kojima, N.
  • Organizer: 2015 Taiwan-Japan Forum on Additive Manufacturing and Bioengineering
  • Place of Presentation: National Formosa University, Yunlin, Taiwan
  • Year and Date: 2015-09-01
  • Int'l Joint Research
• [Presentation] Reconstruction of Bone Marrow Tissue Using Dispersed Cells (2015)
  • Author(s): Sayo, K., Aoki, S. and Kojima, N.
  • Organizer: 2015 Taiwan-Japan Forum on Additive Manufacturing and Bioengineering
  • Place of Presentation: National Formosa University, Yunlin, Taiwan
  • Year and Date: 2015-09-01
  • Int'l Joint Research
• [Presentation] Sugar chain-mediated self-organization of the islet structure in vitro (2015)
  • Author(s): Kojima, N.
  • Organizer: 14th Congress of the Asian Society of Transplantation (CAST) 2015
  • Place of Presentation: Suntec Singapore Convention and Exhibition Centre, Singapore
  • Year and Date: 2015-08-25
  • Int'l Joint Research
• [Presentation] 組織設計技術によるミニチュア臓器のつくりかた (2015)
  • Author(s): 小島伸彦
  • Organizer: 第34回分子病理学研究会
  • Place of Presentation: 兵庫県・神戸ホテルフルーツ・フラワー
  • Year and Date: 2015-07-26
  • Invited
• [Presentation] 骨髄細胞の３次元的再構築 (2015)
  • Author(s): 小島伸彦
  • Organizer: 第22回HAB研究機構学術年会
  • Place of Presentation: 東京都・昭和大学上條講堂
  • Year and Date: 2015-06-26
• [Presentation] 肝スフェロイドはどこまで肝組織に近づけるか？ (2015)
  • Author(s): 小島伸彦
  • Organizer: 第22回肝細胞研究会
  • Place of Presentation: 鳥取県・米子コンベンションセンター
  • Year and Date: 2015-06-04
  • Invited
• [Presentation] 新規手法によるスフェロイドへの細胞外マトリクス充填とその効果 (2015)
  • Author(s): 田尾文哉，小島伸彦
  • Organizer: 第22回肝細胞研究会
  • Place of Presentation: 鳥取県・米子コンベンションセンター
  • Year and Date: 2015-06-04
• [Presentation] Reconstruction of cell polarity in microstructural spheroid (2015)
  • Author(s): Kojima, N., Motoyama, W. and Aoki, S.
  • Organizer: International Forum on MicroManifacturing and Biofabrication 2015
  • Place of Presentation: Toyama International Conference Center, Toyama
  • Year and Date: 2015-05-19
  • Int'l Joint Research
• [Presentation] Induction of hepatic functions in the ECM-loaded spheroids (2015)
  • Author(s): Tao, F., Sato, T., and Kojima, N.
  • Organizer: International Forum on MicroManifacturing and Biofabrication 2015
  • Place of Presentation: Toyama International Conference Center, Toyama
  • Year and Date: 2015-05-19
  • Int'l Joint Research
• [Presentation] Sugar chain mediated autonomous organization of the islets in vitro (2015)
  • Author(s): Kamitori, S., Ozeki, Y., and Kojima, N.
  • Organizer: International Forum on MicroManifacturing and Biofabrication 2015
  • Place of Presentation: Toyama International Conference Center, Toyama
  • Year and Date: 2015-05-19
  • Int'l Joint Research
• [Presentation] Fabrication of bone marrow-like tissue in vitro (2015)
  • Author(s): Sayo, K., Aoki, S., and Kojima, N.
  • Organizer: International Forum on MicroManifacturing and Biofabrication 2015
  • Place of Presentation: Toyama International Conference Center, Toyama
  • Year and Date: 2015-05-19
  • Int'l Joint Research
• [Presentation] 肝細胞の培養に適した微細構造を持つスフェロイドの構築 (2015)
  • Author(s): 小島伸彦，本山和加子，青木茂久
  • Organizer: 第14回日本再生医療学会総会
  • Place of Presentation: パシフィコ横浜
  • Year and Date: 2015-03-20
• [Presentation] 試験管内での膵島構造再構築における糖鎖の関与 (2015)
  • Author(s): 神取紗英，大関泰裕，小島伸彦
  • Organizer: 第14回日本再生医療学会総会
  • Place of Presentation: パシフィコ横浜
  • Year and Date: 2015-03-20
• [Presentation] In vitroにおける骨髄様組織の作製 (2015)
  • Author(s): 佐用かなえ，青木茂久，小島伸彦
  • Organizer: 第14回日本再生医療学会総会
  • Place of Presentation: パシフィコ横浜
  • Year and Date: 2015-03-19
• [Presentation] 組織工学によるハイパー膵島の開発 (2015)
  • Author(s): 小島伸彦
  • Organizer: 細胞アッセイ研究会
  • Place of Presentation: 東京大学生産技術研究所コンベンションホール
  • Year and Date: 2015-01-13
• [Presentation] 膵島を超えた膵島の開発 (2014)
  • Author(s): 小島伸彦
  • Organizer: 第2回細胞凝集研究会
  • Place of Presentation: アクロス福岡
  • Year and Date: 2014-12-06
• [Presentation] In vitro３次元組織におけるマイクロビーズを用いた細胞極性の制御 (2014)
  • Author(s): 本山和加子，青木茂久，小島伸彦
  • Organizer: 第37回日本分子生物学会年会
  • Place of Presentation: パシフィコ横浜
  • Year and Date: 2014-11-27
• [Presentation] 試験管内におけるハイパー膵島の作製 （Fabrication of the hyper-islets in vitro） (2014)
  • Author(s): 小島伸彦
  • Organizer: イノベーション・ジャパン2014－大学見本市
  • Place of Presentation: 東京ビッグサイト
  • Year and Date: 2014-09-11 – 2014-09-12
• [Presentation] Protection from Cell Death in Multicellular Spheroids (2014)
  • Author(s): Motoyama, W., Aoki, S. and Kojima, N.
  • Organizer: 9th World Congress on Alternatives and Animal Use in the Life Sciences
  • Place of Presentation: Hotel Hilton Prague, Prague, Czech Republic
  • Year and Date: 2014-08-25
• [Presentation] 微小環境を再現したミニチュア臓器作製の取り組み (2014)
  • Author(s): 小島伸彦
  • Organizer: 若手研究者の会、病理学、肝臓・糖尿病・内分泌内科学合同特別セミナー
  • Place of Presentation: 佐賀大学医学部・2Fセミナー室2260
  • Year and Date: 2014-08-01
  • Invited
• [Presentation] 人体に設計図はあるのか？ (2014)
  • Author(s): 小島伸彦
  • Organizer: 夢ナビライブ2014
  • Place of Presentation: 東京ビッグサイト
  • Year and Date: 2014-07-12
• [Presentation] 上皮細胞に適した３次元スフェロイド培養法の開発 (2014)
  • Author(s): 本山和加子，青木茂久，小島伸彦
  • Organizer: 第21回肝細胞研究会
  • Place of Presentation: 東京医科歯科大学M&Dタワー
  • Year and Date: 2014-06-27 – 2014-06-28
• [Presentation] Functional Enhancement of Multicellular Spheroid by Microchannel Fabrication (2014)
  • Author(s): Kojima, N.
  • Organizer: TERMIS EU 2014 Chapter Meeting
  • Place of Presentation: Magazzini del Cotone Conference Center, Genova, Italy
  • Year and Date: 2014-06-13
• [Presentation] 内部構造の制御による肝および膵島様スフェロイドの高機能化 (2014)
  • Author(s): 小島伸彦
  • Organizer: 内部構造の制御による肝および膵島様スフェロイドの高機能化
  • Place of Presentation: 東京都千代田区・星陵会館
  • Year and Date: 2014-05-29
  • Invited
• [Presentation] Fabrication of Multicellular Spheroids with Microstructures (2014)
  • Author(s): Kojima, N.
  • Organizer: JST ERATO International Symposium on 3D Tissue Fabrication
  • Place of Presentation: Institute of Industrial Science, The University of Tokyo Cenvention Hall, Tokyo, Japan
  • Year and Date: 2014-05-20
• [Presentation] 微小流路をもつ細胞凝集体を作製する技術とその効果 (2014)
  • Author(s): 小島伸彦
  • Organizer: 第21回HAB研究機構学術年会
  • Place of Presentation: 昭和大学上條講堂
  • Year and Date: 2014-05-16
• [Presentation] 微小環境を再現したミニチュア臓器作製の取り組み (2014)
  • Author(s): 小島伸彦
  • Organizer: 創薬・探索研究所セミナー
  • Place of Presentation: 大阪府豊中市・塩野義製薬株式会・社医薬研究センター
  • Year and Date: 2014-04-23
  • Invited
• [Book] 三次元ティッシュエンジニアリング (2015)
  • Author(s): 小島伸彦（分担）
  • Total Pages: 400
  • Publisher: エヌ・ティー・エス
• [Remarks] 再生生物学研究室（小島伸彦研究室）
  • URL: http://regenbio.sci.yokohama-cu.ac.jp
• [Patent(Industrial Property Rights)] 高分子を細胞と共に凝集させる技術 (2015)
  • Inventor(s): 小島伸彦
  • Industrial Property Rights Holder: 公立大学法人横浜市立大学
  • Industrial Property Rights Type: 特許
  • Filing Date: 2015-05-14
• [Patent(Industrial Property Rights)] 骨髄細胞凝集体の作製方法 (2015)
  • Inventor(s): 小島伸彦
  • Industrial Property Rights Holder: 公立大学法人横浜市立大学
  • Industrial Property Rights Type: 特許
  • Filing Date: 2015-01-28