Visualization of functional hepatic cells derived from human iPS cells.

• Project/Area Number: 26670585
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: General surgery
• Research Institution: Yokohama City University
• Principal Investigator: SEKINE Keisuke 横浜市立大学, 医学部, 助教 (00323569)
• Co-Investigator(Kenkyū-buntansha): TAKEBE Takanori 横浜市立大学, 医学部, 准教授 (20612625)
• Project Period (FY): 2014-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥3,640,000 (Direct Cost: ¥2,800,000、Indirect Cost: ¥840,000); Fiscal Year 2015: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2014: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
• Keywords: 再生医療 / 細胞・組織 / 再生医学 / 幹細胞 / 肝疾患

Outline of Final Research Achievements

In this study, we evaluated feeder-free iPS cell culture condition for producing a knock- iPS cells, and evaluated optimal transfection conditions.Next, we established the knock-in iPS cells constitutively express a fluorescent protein. It was confirmed that the knock-in iPS cell clone expressed fluorescent protein even after differentiation induction. Furthermore, we succeeded to establish a knock-in iPS cells targeted for the hepatic differentiation marker gene. The established iPS cell could be useful to determine the optimal timing of hepatic cells for producing IPSC-liver bud toward clinical applications.

Research Products

• [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 Issue: 4 Pages: 1205-1207
  • DOI: 10.1016/j.transproceed.2014.02.003
  • 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 Issue: 8 Pages: 0-2
  • DOI: 10.1371/journal.pone.0104776
  • Peer Reviewed / Open Access
• [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] Efficient hepatic differentiation of human induced pluripotent stem cells in a three-dimensional microscale culture. (2014)
  • Author(s): Zhang RR, Takebe T, Miyazaki L, Takayama M, Koike H, Kimura M, Enomura M, Zheng YW, Sekine K, Taniguchi H.
  • Journal Title: Methods Mol Biol. Volume: 1210 Pages: 131-141
  • DOI: 10.1007/978-1-4939-1435-7_10
  • ISBN: 9781493914340, 9781493914357
  • Peer Reviewed
• [Journal Article] The evidence of sophisticatedly heterogeneous population of human umbilical vein endothelial cells (2014)
  • Author(s): Zheng YW, Nie YZ, Tsuchida T, Zhang RR, Aoki K, Sekine K, Ogawa M, Takebe T, Ueno Y, Sakakibara H, Hirahara F, Taniguchi H
  • Journal Title: Transplant Proc Volume: 46 Issue: 4 Pages: 1251-3
  • DOI: 10.1016/j.transproceed.2013.11.077
  • Peer Reviewed / Open Access
• [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(4) Issue: 4 Pages: 1201-4
  • DOI: 10.1016/j.transproceed.2013.12.021
  • 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 Issue: 4 Pages: 1191-3
  • DOI: 10.1016/j.transproceed.2013.12.026
  • Peer Reviewed / Open Access
• [Journal Article] High-resolution intravital imaging for monitoring the transplanted islets 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 Issue: 4 Pages: 1166-1168
  • DOI: 10.1016/j.transproceed.2013.11.089
  • 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, and Taniguchi H
  • Journal Title: Stem Cells Dev Volume: (In press) Issue: 18 Pages: 2237-2249
  • DOI: 10.1089/scd.2013.0577
  • Peer Reviewed / Open Access
• [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, Nakata S, Ouchi R, Obana Y, Sekine K, Zheng YW, Takebe T, Isono K, Koseki H, Taniguchi H
  • Journal Title: Hepatology Volume: 印刷中 Issue: 1 Pages: 323-333
  • DOI: 10.1002/hep.27046
  • Peer Reviewed / Open Access
• [Journal Article] Generation of a vascularized and functional human liver from an iPSC-derived organ bud transplant. (2014)
  • Author(s): Takanori Takebe*, Ran-Ran Zhang, Hiroyuki Koike, Masaki Kimura, Emi Yoshizawa, Masahiro Enomura, Keisuke Sekine, Hideki Taniguchi*(*: Correspondence)
  • Journal Title: Nature Protocols Volume: 9(2) Issue: 2 Pages: 396-409
  • DOI: 10.1038/nprot.2014.020
  • Peer Reviewed
• [Presentation] シングルセルRNAシークエンスによるiPS細胞由来肝細胞のポピュレーション解析 (2016)
  • Author(s): 関根圭輔、J. Gray Camp、Barbara Treutlein、武部 貴則、谷口 英樹
  • Organizer: 第15回日本再生医療学会総会
  • Place of Presentation: 大阪国際会議場(大阪府大阪市)
  • Year and Date: 2016-03-17
• [Presentation] Generation of functional human liver from pluripotent stem cell (2016)
  • Author(s): Keisuke Sekine, Takanori Takebe, Hideki Taniguchi
  • Organizer: German Association for the Study of the Liver (GASL) workshop
  • Place of Presentation: Dusseldorf(ドイツ)
  • Year and Date: 2016-01-21
  • Int'l Joint Research / Invited
• [Presentation] iPS細胞を用いた代謝性臓器の再生医療 (2015)
  • Author(s): 関根圭輔
  • Organizer: 第30回日本整形外科学会基礎学術総会
  • Place of Presentation: 富山市民プラザ(富山県富山市)
  • Year and Date: 2015-10-22
  • Invited
• [Presentation] Pdx1+ cells behave as pancreatic cancer stem cells with high tumorigenicity and metastatic potential (2015)
  • Author(s): Ryo Okuda, Keisuke Sekine, Lue Sun , Kaori Hamanaka, Hideki Taniguchi
  • Organizer: 第74回日本癌学会学術総会
  • Place of Presentation: 名古屋国際会議場(愛知県名古屋市)
  • Year and Date: 2015-10-08
• [Presentation] Metastatic tropism of pancreatic cancer stem cells to a specific distant organ. (2015)
  • Author(s): Ryo Okuda, Keisuke Sekine, Kaori Hamanaka, Hideki Taniguchi.
  • Organizer: The Biology of Cancer: Microenvironment, Metastasis & Therapeutics
  • Place of Presentation: New York (米国)
  • Year and Date: 2015-05-12
  • Int'l Joint Research
• [Presentation] iPS細胞を用いた代謝性臓器の再生医療 (2015)
  • Author(s): 武部貴則、関根圭輔、谷口英樹
  • Organizer: 第14回日本再生医療学会総会
  • Place of Presentation: パシフィコ横浜（神奈川県）
  • Year and Date: 2015-03-19 – 2015-03-21
  • Invited
• [Presentation] A novel system with human liver features for screening anti-hepatitis B virus therapeutics (2015)
  • Author(s): Nie YZ,Zheng YW,kZhang RR,Tsuchida T,Ueno Y、Sekine K,Takebe T,Taniguchi H
  • Organizer: 第14回日本再生医療学会総会
  • Place of Presentation: パシフィコ横浜（神奈川県）
  • Year and Date: 2015-03-19 – 2015-03-21
• [Presentation] 膵癌転移における微小環境と癌幹細胞の相互作用の解析 (2015)
  • Author(s): 濱中香織、奥田諒、関根圭輔、谷口英樹
  • Organizer: 第14回日本再生医療学会総会
  • Place of Presentation: パシフィコ横浜（神奈川県）
  • Year and Date: 2015-03-19 – 2015-03-21
• [Presentation] 膵癌幹細胞の転移における特定臓器への指向性 (2015)
  • Author(s): 泉陽彦、竹歳卓人、関根圭輔、奥田諒、濱中香織、谷口英樹
  • Organizer: 第14回日本再生医療学会総会
  • Place of Presentation: パシフィコ横浜（神奈川県）
  • Year and Date: 2015-03-19 – 2015-03-21
• [Presentation] 遺伝子改変モデル動物を用いた膵癌幹細胞の機能解析 (2014)
  • Author(s): 関根圭輔、奥田諒、久松大介、濱中香織、谷口英樹
  • 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 – 2014-11-29
  • Invited
• [Presentation] Identification and characterization of Pdx1+ pancreatic cancer stem cell during pancreatic tumorigenesis (2014)
  • Author(s): 関根圭輔、奥田諒、久松大介、谷口英樹
  • Organizer: The 73rd Annual Meeting of the Japanese Cancer Association.
  • Place of Presentation: パシフィコ横浜（神奈川県）
  • Year and Date: 2014-09-25 – 2014-09-27
• [Presentation] スフィア形成能を有する膵癌幹細胞の解析 (2014)
  • Author(s): 久松大介、奥田諒、関根圭輔、濵中香織、上野康晴、谷口英樹
  • Organizer: The 73rd Annual Meeting of the Japanese Cancer Association.
  • Place of Presentation: パシフィコ横浜（神奈川県）
  • Year and Date: 2014-09-25 – 2014-09-27
• [Presentation] Identification of pancreatic cancer stem cell lineage during pancreatic tumorigenesis (2014)
  • Author(s): 奥田諒、関根圭輔、久松大介、孫略、濵中香織、谷口英樹
  • Organizer: The 73rd Annual Meeting of the Japanese Cancer Association.
  • Place of Presentation: パシフィコ横浜（神奈川県）
  • Year and Date: 2014-09-25 – 2014-09-27
• [Patent(Industrial Property Rights)] 癌微小環境を再現できる癌組織の再構築法 (2016)
  • Inventor(s): 谷口英樹、関根圭輔、上野康晴、奥田諒、武部貴則
  • Industrial Property Rights Holder: 谷口英樹、関根圭輔、上野康晴、奥田諒、武部貴則
  • Industrial Property Rights Type: 特許
  • Filing Date: 2016-03-16
• [Patent(Industrial Property Rights)] 血液細胞を用いた組織・臓器の作製方法 (2015)
  • Inventor(s): 谷口英樹、武部貴則、関根圭輔、鄭允文、聶運中
  • Industrial Property Rights Holder: 谷口英樹、武部貴則、関根圭輔、鄭允文、聶運中
  • Industrial Property Rights Type: 特許
  • Filing Date: 2015-12-24