Acute liver failure treatment using rapid fabricated engineered hepatocyte sheets

• Project/Area Number: 26461916
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: General surgery
• Research Institution: Nagasaki University
• Principal Investigator: EGUCHI Susumu 長崎大学, 医歯薬学総合研究科(医学系), 教授 (80404218)
• Co-Investigator(Kenkyū-buntansha): 山之内 孝彰 長崎大学, 医歯薬学総合研究科(医学系), 講師 (10448508) ; 堺 裕輔 長崎大学, 医歯薬学総合研究科(医学系), 助教 (10608904)
• Research Collaborator: KOIKE Makiko ; HASEGAWA Hideko ; OHASHI Kazuo ; TAKATSUKI Mitsuhisa ; HIDAKA Masaaki ; SOYAMA Akihiko
• Project Period (FY): 2014-04-01 – 2018-03-31
• Project Status: Completed (Fiscal Year 2017)
• Budget Amount: ¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000); Fiscal Year 2016: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2015: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2014: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
• Keywords: 急性肝不全 / 肝細胞 / 細胞シート / 再生医療

Outline of Final Research Achievements

It is desired to establish a new technology that can support complicated and diverse liver specific-functions that enables hasty treatment for acute liver failure. In this study, we aimed to establish rapid production technology of engineered hepatocyte sheet which makes it possible to treat acute liver failure.
Primary human hepatocytes were isolated from non-tumor area of surgically excised liver tissue. The human fibroblasts (TIG -118 cells) at a density of 1500000 cells/dish or more formed stably confluent onto FBS-coated 35-mm diameter temperature responsive culture dish. Primary human hepatocytes were quickly adhered onto TIG-118 cell layer. The engineered hepatocyte sheets could be harvested as a cell sheet within a total of 4 hours.

Research Products

• [Journal Article] Spontaneous hepatocyte migration towards an endothelial cell tube network (2018)
  • Author(s): Baimakhanov Zhassulan、Sakai Yusuke、Yamanouchi Kosho、Hidaka Masaaki、Soyama Akihiko、Takatsuki Mitsuhisa、Eguchi Susumu
  • Journal Title: Journal of Tissue Engineering and Regenerative Medicine Volume: 12 Issue: 3
  • DOI: 10.1002/term.2577
  • Peer Reviewed
• [Journal Article] Controlled cell morphology and liver-specific function of engineered primary hepatocytes by fibroblast layer cell densities (2018)
  • Author(s): Sakai Yusuke、Koike Makiko、Kawahara Daisuke、Hasegawa Hideko、Murai Tomomi、Yamanouchi Kosho、Soyama Akihiko、Hidaka Masaaki、Takatsuki Mitsuhisa、Fujita Fumihiko、Kuroki Tamotsu、Eguchi Susumu
  • Journal Title: Journal of Bioscience and Bioengineering Volume: 印刷中 Issue: 2 Pages: 249-257
  • DOI: 10.1016/j.jbiosc.2018.02.006
  • NAID: 40021645454
  • Peer Reviewed
• [Journal Article] Tissue Repair Promotion and Functional Tissue Fabrication of Digestive Organ by Cell Sheets (2018)
  • Author(s): 堺 裕輔、江口 晋
  • Journal Title: MEMBRANE Volume: 43 Issue: 2 Pages: 34-39
  • DOI: 10.5360/membrane.43.34
  • NAID: 130006638816
  • ISSN: 0385-1036, 1884-6440
• [Journal Article] In vivo construction of liver tissue by implantation of a hepatic non-parenchymal/adipose-derived stem cell sheet (2017)
  • Author(s): Mio Fujii, Kosho Yamanouchi, Yusuke Sakai, Zhassulan Baimakhanov, Izumi Yamaguchi, Akihiko Soyama, Masaaki Hidaka, Mitsuhisa Takatsuki, Tamotsu Kuroki, Susumu Eguchi
  • Journal Title: Journal of Tissue Engineering and Regenerative Medicine Volume: 印刷中 Issue: 1 Pages: e287-e295
  • DOI: 10.1002/term.2424
  • Peer Reviewed
• [Journal Article] 細胞を用いた肝再生医療の最前線 (2017)
  • Author(s): 岡田 尚子、堺 裕輔、江口 晋
  • Journal Title: PHARMSTAGE Volume: 17 Pages: 10-16
• [Journal Article] Toyocamycin attenuates free fatty acid-induced hepatic steatosis and apoptosis in cultured hepatocytes and ameliorates nonalcoholic fatty liver disease in mice (2017)
  • Author(s): Ikuko Takahara, Yuko Akazawa, Maiko Tabuchi, Katsuya Matsuda, Hisamitsu Miyaaki, Youko Kido, Yasuko Kanda, Naota Taura, Ken Ohnita, Fuminao Takeshima, Yusuke Sakai, Susumu Egushi, Masahiro Nakashima, Kazuhiko Nakao
  • Journal Title: PLoS One Volume: 12 Issue: 3 Pages: e0170591-e0170591
  • DOI: 10.1371/journal.pone.0170591
  • NAID: 120006987538
  • Peer Reviewed / Open Access
• [Journal Article] Equal distribution of MSCs after hepatic ischemia reperfusion injury (2016)
  • Author(s): Askhat Isbambetov, Zhassulan Baimakhanov, Akihiko Soyama, Masaaki Hidaka, Yusuke Sakai, Mitsuhisa Takatsuki, Tamotsu Kuroki, Susumu Eguchi
  • Journal Title: Journal of Surgical Research Volume: 203 Issue: 2 Pages: 360-367
  • DOI: 10.1016/j.jss.2016.03.057
  • Peer Reviewed
• [Journal Article] Up-regulated extracellular matrix components and inflammatory chemokines may impair the regeneration of cholestatic liver (2016)
  • Author(s): Shuai Zhang, Tao-Sheng Li, Akihiko Soyama, Takayuki Tanaka, Chen Yan, Yusuke Sakai, Masaaki Hidaka, Ayaka Kinoshita, Koji Natsuda, Mio Fujii, Tota Kugiyama, Zhassulan Baimakhanov, Tamotsu Kuroki, Weili Gu, Susumu Eguchi
  • Journal Title: Scientific Reports Volume: 6 Issue: 1 Pages: 26540-26540
  • DOI: 10.1038/srep26540
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Vascularized subcutaneous human liver tissue from engineered hepatocyte/fibroblast sheets in mice (2015)
  • Author(s): Yusuke Sakai, Kosho Yamanouchi, Kazuo Ohashi, Makiko Koike, Rie Utoh, Hideko Hasegawa, Izumi Muraoka, Takashi Suematsu, Akihiko Soyama, Masaaki Hidaka, Mitsuhisa Takatsuki, Tamotsu Kuroki, Susumu Eguchi
  • Journal Title: Biomaterials Volume: 65 Pages: 66-75
  • DOI: 10.1016/j.biomaterials.2015.06.046
  • NAID: 120006986875
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Efficacy of multi-layered hepatocyte sheet transplantation for radiation-induced liver damage and partial hepatectomy in a rat model (2015)
  • Author(s): Zhassulan Baimakhanov, Kosho Yamanouchi, Yusuke Sakai, Makiko Koike, Akihiko Soyama, Masaaki Hidaka, Mitsuhisa Takatsuki, Fumihiko Fujita, Kengo Kanetaka, Tamotsu Kuroki, Susumu Eguchi
  • Journal Title: Cell Transplantation Volume: 25 Issue: 3 Pages: 549-558
  • DOI: 10.3727/096368915x688669
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Rapid production of engineered human primary hepatocyte/fibroblast sheets (2015)
  • Author(s): Yusuke Sakai, Makiko Koike, Akihiko Soyama, Masaaki Hidaka, Tamotsu Kuroki, Susumu Eguchi
  • Journal Title: Data in Brief Volume: 5 Pages: 498-501
  • DOI: 10.1016/j.dib.2015.09.044
  • NAID: 120006986862
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Efficacy of multi-layered hepatocyte sheet transplantation for radiation-induced liver damage and partial hepatectomy in a rat model (2015)
  • Author(s): Zhassulan Baimakhanov, Kosho Yamanouchi, Yusuke Sakai, Akihiko Soyama, Masaaki Hidaka, Mitsuhisa Takatsuki, Fumihiko Fujita, Kengo Kanetaka, Tamotsu Kuroki, Susumu Eguchi
  • Journal Title: Journal of the American College of Surgeons Volume: 221
  • Peer Reviewed / Open Access
• [Journal Article] 組織工学による低侵襲な肝疾患治療 (2015)
  • Author(s): 堺裕輔、江口晋
  • Journal Title: 細胞 Volume: 47 Pages: 18-21
• [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] 細胞シート工学 (2014)
  • Author(s): 小林慎一朗, 岡野光夫, 江口晋
  • Journal Title: 消化器外科 Volume: 37 Pages: 1321-1332
• [Presentation] 効率的・移植可能な肝胆膵領域での再生医療 (2018)
  • Author(s): 江口晋、黄宇、堺裕輔
  • Organizer: 第17回日本再生医療学会総会
  • Invited
• [Presentation] 術後急性肝不全予防のための迅速な肝細胞複合シート作製 (2017)
  • Author(s): 堺裕輔、小池真章子、曽山明彦、日高匡章、高槻光寿、江口晋
  • Organizer: 第16回日本再生医療学会総会
  • Place of Presentation: 仙台国際センター（宮城県、仙台市）
  • Year and Date: 2017-03-07
• [Presentation] 細胞シート工学を利用した血管網を有する皮下培養型ヒト肝組織作製 (2017)
  • Author(s): 堺裕輔、高槻光寿、江口晋
  • Organizer: 第103回日本消化器病学会総会
• [Presentation] 肝特異的な特徴を有するヒト肝組織の皮下構築と機能発現解析 (2016)
  • 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、江口晋
  • Organizer: 第23回肝細胞研究会
  • Place of Presentation: 大阪大学中之島センター（大阪府、大阪市）
  • Year and Date: 2016-07-07
• [Presentation] 安定した肝細胞複合シート作製のための線維芽細胞培養条件の検討 (2016)
  • Author(s): 小池真章子、堺裕輔、川原大輔、高槻光寿、江口晋
  • Organizer: 第23回肝細胞研究会
  • Place of Presentation: 大阪大学中之島センター（大阪府、大阪市）
  • Year and Date: 2016-07-07
• [Presentation] 皮下性ヒト肝組織構築に伴う血管網と肝特異機能発現の評価 (2016)
  • Author(s): 堺裕輔、小池真章子、山之内孝彰、曽山明彦、日高匡章、黒木保、江口晋
  • Organizer: 第15回日本再生医療学会総会
  • Place of Presentation: 大阪国際会議場（大阪）
  • Year and Date: 2016-03-17
• [Presentation] Efficacy of multi-layered hepatocyte sheet transplantation for radiation-induced liver damage and partial hepatectomy in a rat model (2015)
  • Author(s): Zhassulan Baimakhanov, Kosho Yamanouchi, Yusuke Sakai, Akihiko Soyama, Masaaki Hidaka, Mitsuhisa Takatsuki, Fumihiko Fujita, Kengo Kanetaka, Tamotsu Kuroki, Susumu Eguchi
  • Organizer: Clinical Congress 2015, American College of Surgeons
  • Place of Presentation: Chicago, IL, USA
  • Year and Date: 2015-10-04
  • Int'l Joint Research
• [Presentation] 皮下性ヒト肝組織構築による肝疾患治療 (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、黒木保、江口晋
  • Organizer: 第22回肝細胞研究会
  • Place of Presentation: 米子コンベンションセンター（鳥取）
  • Year and Date: 2015-06-04
• [Presentation] 支持細胞密度による初代肝細胞の形態および機能発現制御 (2015)
  • Author(s): 小池真章子、堺裕輔、川原大輔、黒木保、江口晋
  • Organizer: 第22回肝細胞研究会
  • Place of Presentation: 米子コンベンションセンター（鳥取）
  • Year and Date: 2015-06-04
• [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] 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, Susumu Eguchi
  • Organizer: ASGCT 18th Annual Meeting
  • Place of Presentation: New Orleans, LA, USA
  • Year and Date: 2015-05-13
  • Int'l Joint Research
• [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
  • Invited
• [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
• [Book] 再生医療用語ハンドブック (2015)
  • Author(s): 江口晋 共著
  • Total Pages: 5
  • Publisher: 株式会社メディカルトリビューン