New method for establishing self-renewing megakaryocyte cell lines using iPS cells

• Project/Area Number: 18K18365
• Research Category: Grant-in-Aid for Early-Career Scientists
• Allocation Type: Multi-year Fund
• Review Section: Basic Section 90110:Biomedical engineering-related
• Research Institution: Kyoto University
• Principal Investigator: Nakamura Sou 京都大学, iPS細胞研究所, 特定拠点助教 (50769833)
• Project Period (FY): 2018-04-01 – 2022-03-31
• Project Status: Completed (Fiscal Year 2021)
• Budget Amount: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000); Fiscal Year 2021: ¥780,000 (Direct Cost: ¥600,000、Indirect Cost: ¥180,000); Fiscal Year 2020: ¥780,000 (Direct Cost: ¥600,000、Indirect Cost: ¥180,000); Fiscal Year 2019: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2018: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
• Keywords: 巨核球 / iPS細胞 / imMKCL / iPS / 血小板 / 自己複製 / c-MYC / 巨核球分化 / 再生医療

Outline of Final Research Achievements

RNA-seq analysis of several megakaryocyte cell lines displaying different proliferative capabilities enabled to identify the responsible genes that inhibit self-renewal ability in megakaryocyte cell lines. Furthermore, alternative vector system was applied to generate megakaryocyte cell lines, whereby epigenetic modifications were silenced at the stage of hematopoietic cell differentiation from pluripotent stem cells. Such improvements allowed us to generate megakaryocyte cell lines from several human iPS cell clones, contributing to high efficiency of megakaryocyte cell line induction.

Academic Significance and Societal Importance of the Research Achievements

同種免疫性の血小板輸血不応症を発症する患者では、HLA/HPAが適合するドナーの確保が必要であるが、稀な型の場合や緊急時は供給不足が危惧される。解決策として、申請者らは、iPS細胞から分化誘導した血小板前駆細胞、巨核球を細胞株化し、産業化可能な血小板製造法を確立したが、安定増殖株の樹立効率が低いことが課題となっていた。本研究で明らかとなった巨核球細胞株自己複製抑制機構を解除する方法を新規ベクターシステムに新たに組み込むことで、個性の異なる複数のiPS細胞クローン由来不死化巨核球の作製効率を飛躍的に改善することができるため、HLA/HPA一致血小板製剤供給源の問題点を解決する可能性が提示できた。

Research Products

• [Journal Article] Extracellular laminin regulates hematopoietic potential of pluripotent stem cells through integrin β1-ILK-β-catenin-JUN axis (2021)
  • Author(s): Yuzuriha Akinori、Nakamura Sou、Sugimoto Naoshi、Kihara Shunsuke、Nakagawa Masato、Yamamoto Takuya、Sekiguchi Kiyotoshi、Eto Koji
  • Journal Title: Stem Cell Research Volume: 53 Pages: 102287-102287
  • DOI: 10.1016/j.scr.2021.102287
  • NAID: 120007006753
  • Peer Reviewed / Open Access
• [Journal Article] Microfluidic Bioreactor Made of Cyclo-Olefin Polymer for Observing On-Chip Platelet Production (2021)
  • Author(s): Kumon Hiroki、Sakuma Shinya、Nakamura Sou、Maruyama Hisataka、Eto Koji、Arai Fumihito
  • Journal Title: Micromachines Volume: 12 Issue: 10 Pages: 1253-1253
  • DOI: 10.3390/mi12101253
  • Peer Reviewed / Open Access
• [Journal Article] Silencing of p53 and CDKN1A establishes sustainable immortalized megakaryocyte progenitor cells from human iPSCs (2021)
  • Author(s): Sone Masamitsu、Nakamura Sou、Umeda Sachiko、Ginya Harumi、Oshima Motohiko、Kanashiro Maria Alejandra、Paul Sudip Kumar、Hashimoto Kanae、Nakamura Emiri、Harada Yasuo、Tsujimura Kyoko、Saraya Atsunori、Yamaguchi Tomoyuki、Sugimoto Naoshi、Sawaguchi Akira、Iwama Atsushi、Eto Koji、Takayama Naoya
  • Journal Title: Stem Cell Reports Volume: 16 Issue: 12 Pages: 2861-2870
  • DOI: 10.1016/j.stemcr.2021.11.001
  • NAID: 120007177810
  • Peer Reviewed / Open Access
• [Journal Article] Development of platelet replacement therapy using human induced pluripotent stem cells (2021)
  • Author(s): Nakamura Sou、Sugimoto Naoshi、Eto Koji
  • Journal Title: Development, Growth & Differentiation Volume: 63 Issue: 2 Pages: 178-186
  • DOI: 10.1111/dgd.12711
  • Peer Reviewed / Open Access
• [Journal Article] Ex vivo generation of platelet products from human iPS cells. (2020)
  • Author(s): Nakamura S, Sugimoto N, Eto K.
  • Journal Title: Inflamm Regen. Volume: 40 Issue: 1 Pages: 10-10
  • DOI: 10.1186/s41232-020-00139-2
  • Peer Reviewed / Open Access
• [Journal Article] Novel physico-chemical regulation mechanism enables the production of 100 billion platelets (2020)
  • Author(s): Nakamura S, Eto K.
  • Journal Title: Rinsho Ketsueki Volume: 61 Issue: 6 Pages: 628-633
  • DOI: 10.11406/rinketsu.61.628
  • NAID: 130007869141
  • ISSN: 0485-1439, 1882-0824
  • Peer Reviewed
• [Presentation] iPS細胞を用いた血小板の作製 (2020)
  • Author(s): 中村 壮
  • Organizer: 第14回日本血栓止血学会学術標準化委員会シンポジウム
  • Invited
• [Patent(Industrial Property Rights)] 国立大学法人千葉大学,国立大学法人京都大学 (2020)
  • Inventor(s): 髙山直也,江藤浩之,曽根正光,中村壮
  • Industrial Property Rights Holder: 髙山直也,江藤浩之,曽根正光,中村壮
  • Industrial Property Rights Type: 特許
  • Filing Date: 2020
  • Overseas