Construction of vascularized ectopic liver tissue from engineered human hepatocyte sheet

• Project/Area Number: 25861161
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: General surgery
• Research Institution: Nagasaki University
• Principal Investigator: SAKAI Yusuke 長崎大学, 医歯薬学総合研究科(医学系), 助教 (10608904)
• Project Period (FY): 2013-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000); Fiscal Year 2014: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000); Fiscal Year 2013: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
• Keywords: 肝細胞 / 細胞シート / 再生医療 / 血管新生

Outline of Final Research Achievements

Subcutaneous liver tissue engineering is an attractive and minimally invasive approach as liver regenerative medicine. In this study, we describe a subcutaneous human liver construction allowing for rapidly vascularized grafts by transplanting engineered hepatocyte/fibroblast sheets (EHFSs).
EHFSs were thicker than hepatocyte-only sheets (HSs), this result could aide in easy manipulation during transplantation. The EHFSs showed superior expression levels of vascularization-associated growth factors (VEGF, TGFb1, and HGF) in vitro. EHFSs were transplanted subcutaneously into immunodeficient mice. The EHFSs formed vascularized subcutaneous human liver tissues (VSLTs) by rapidly neo-vascularization under mouse skin. Furthermore, subacute hepatic failure model mice with VSLT were alive at least 7 weeks after liver damage. This approach could prove valuable for establishing novel cell therapies for liver diseases.

Research Products

• [Journal Article] Detachably assembled microfluidic device for perfusion culture and post-culture analysis of a spheroid array (2014)
  • Author(s): Yusuke Sakai, Koji Hattori, Fumiki Yanagawa, Shinji Sugiura, Toshiyuki Kanamori, and Kohji Nakazawa
  • Journal Title: Biotechnology Journal Volume: 9 Issue: 7 Pages: 971-979
  • DOI: 10.1002/biot.201300559
  • Peer Reviewed
• [Journal Article] Micropatterned culture of HepG2 spheroids using microwell chip with honeycomb-patterned polymer film (2014)
  • Author(s): Hidekazu Yamazaki, Shun Goto, Koju Ito, Souichi Kohashi, Yuki Goto, Yukiko Yoshiura, Yusuke Sakai, Hiroshi Yabu, Masatsugu Shimomura, and Kohji Nakazawa
  • Journal Title: Journal of Bioscience and Bioengineering Volume: 118 Issue: 4 Pages: 455-460
  • DOI: 10.1016/j.jbiosc.2014.03.006
  • NAID: 110009865430
  • Peer Reviewed
• [Journal Article] Rapid fabrication of engineered liver tissue using novel fibroblast system (2014)
  • Author(s): 1.Yusuke Sakai, Makiko Koike, Hideko Hasegawa, Kosho Yamanouchi, Akihiko Soyama, Masaaki Hidaka, Mitsuhisa Takatsuki, Tamotsu Kuroki, Teruo Okano, and Susumu Eguchi
  • Journal Title: ALTEX Proceedings Volume: 3 Pages: 44-44
  • Peer Reviewed / Open Access
• [Journal Article] Rapid fabricating technique for multi-layered human hepatic cell sheets by forceful contraction of the fibroblast monolayer (2013)
  • Author(s): Yusuke Sakai, Makiko Koike, Hideko Hasegawa, Kosho Yamanouchi, Akihiko Soyama, Mitsuhisa Takatsuki, Tamotsu Kuroki, Kazuo Ohashi, Teruo Okano, and Susumu Eguchi
  • Journal Title: PLoS One Volume: 8 (7) Issue: 7 Pages: e70970-e70970
  • DOI: 10.1371/journal.pone.0070970
  • NAID: 120006986139
  • Peer Reviewed
• [Journal Article] Construction of Liver Tissue in vivo with Preparative Partial Hepatic Irradiation and Growth Stimulus: Investigations of Less Invasive Techniques and Progenitor Cells (2013)
  • Author(s): Kensuke Miyazaki, Kosho Yamanouchi, Yusuke Sakai, Izumi Yamaguchi, Mitsuhisa Takatsuki, Tamotsu Kuroki, Chandan Guha, and Susumu Eguchi
  • Journal Title: Journal of Surgical Research Volume: 185 (2) Issue: 2 Pages: 889-895
  • DOI: 10.1016/j.jss.2013.06.016
  • NAID: 120006986772
  • Peer Reviewed
• [Presentation] Reconstruction of Vascularized Subcutaneous Liver Tissue for Minimally Invasive Cell Therapy (2015)
  • Author(s): Yusuke Sakai, Makiko Koike, Kosho Yamanouchi, Akihiko Soyama, Masaaki Hidaka, Mitsuhisa Takatsuki, Tamotsu Kuroki, and Susumu Eguchi
  • Organizer: ASGCT 18th Annual Meeting
  • Place of Presentation: New Orleans, LA, USA
  • Year and Date: 2015-05-13 – 2015-05-16
• [Presentation] 細胞シート工学を利用した血管誘導ヒト肝組織の迅速な自己構築 (2015)
  • Author(s): 堺裕輔、小池真章子、長谷川英子、山之内孝彰、曽山明彦、日高匡章、高槻光寿、黒木保、江口晋
  • Organizer: 第115回日本外科学会定期学術集会
  • Place of Presentation: 名古屋国際会議場（名古屋）
  • Year and Date: 2015-04-16 – 2015-04-18
• [Presentation] Rapid fabrication of engineered liver tissue using novel fibroblast system (2014)
  • Author(s): Yusuke Sakai, Makiko Koike, Hideko Hasegawa, Kosho Yamanouchi, Akihiko Soyama, Masaaki Hidaka, Mitsuhisa Takatsuki, Tamotsu Kuroki, Teruo Okano, and Susumu Eguchi
  • Organizer: 9th World Congress on Alternatives and Animal Use in the Life Sciences (WC9)
  • Place of Presentation: Prague, Czech Republic
  • Year and Date: 2014-08-24 – 2014-08-28
• [Presentation] 異所性肝組織構築のための再生医療デバイスの創成
  • Author(s): 堺裕輔、小池真章子、山之内孝彰、曽山明彦、高槻光寿、黒木保、鵜頭理恵、大橋一夫、岡野光夫、江口晋
  • Organizer: 第68回日本消化器外科学会総会
  • Place of Presentation: 宮崎（シーガイアコンベンションセンター）
• [Presentation] ヒト初代肝細胞/線維芽細胞複合シートが創造する第二の肝臓
  • Author(s): 堺裕輔、高槻光寿、江口晋
  • Organizer: JDDW2013
  • Place of Presentation: 東京（グランドプリンスホテル新高輪）
• [Presentation] 肝移植の代替または再生補助療法としてのヒト肝細胞シートを用いた再生医療デバイスの開発
  • Author(s): 高槻光寿、堺裕輔、江口晋
  • Organizer: 第40回日本肝臓学会西部会
  • Place of Presentation: 岐阜（長良川国際会議場）
• [Presentation] ヒト初代肝細胞複合シート移植における支持細胞の種類の効果
  • Author(s): 堺裕輔、小池真章子、長谷川英子、山之内孝彰、日高匡章、曽山明彦、高槻光寿、黒木保、岡野光夫、江口晋
  • Organizer: 第13回日本再生医療学会総会
  • Place of Presentation: 京都（国立京都国際会館）
• [Presentation] 肝細胞／各種支持細胞共培養シートの構造と機能発現の評価
  • Author(s): 小池真章子、堺裕輔、黒木保、江口晋
  • Organizer: 第13回日本再生医療学会総会
  • Place of Presentation: 京都（国立京都国際会館）
• [Book] 再生医療用語ハンドブック： 肝細胞移植 (2015)
  • Author(s): 堺裕輔、江口晋
  • Total Pages: 1
  • Publisher: 株式会社メディカルトリビューン