Establishment of new human pluripotent stem cell with unbiased differentiation potency.

• Project/Area Number: 18K14714
• Research Category: Grant-in-Aid for Early-Career Scientists
• Allocation Type: Multi-year Fund
• Review Section: Basic Section 44020:Developmental biology-related
• Research Institution: Kyoto University
• Principal Investigator: Nakamura Tomonori 京都大学, 白眉センター, 特定准教授 (90648429)
• Project Period (FY): 2018-04-01 – 2021-03-31
• Project Status: Completed (Fiscal Year 2020)
• Budget Amount: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000); Fiscal Year 2020: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000); Fiscal Year 2019: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2018: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
• Keywords: カニクイザル / 多能性幹細胞 / scRNA-seq / 霊長類 / 高次元空間ノイズ / 次元の呪い / 分化偏向性

Outline of Final Research Achievements

Pluripotent stem cells (PSCs) are cells that can proliferate indefinitely while retaining the ability to contribute to whole adult cells. In this study, we attempted to establish a new culture system that eliminates the differentiation bias seen in human PSCs by mimicking the environment in vivo. Since the target in vivo pluripotent cells correspond immediately after implantation, we aimed to analyze the traits of cynomolgus monkey implantation embryos using scRNA-seq, examine the culture conditions using the monkey PSC, and reproduce them in human PSC. In this process, we have established a new robust and stable culture system for monkey PSC, which has been unstable, and surprisingly discovered a universal problem in scRNA-seq analysis, and found a solution.

Academic Significance and Societal Importance of the Research Achievements

ヒトiPSCが樹立されて以来、モデル動物としての非ヒト霊長類が見直されてきている。また、霊長類に関わらず昨今の分子生物学ではscRNA-seqを筆頭に単一細胞レベルの研究領域に突入している。これらの状況を鑑みると本研究成果はこれからの時代の基礎となる知見や技術になり、学術的意義は高いと考える。今後本研究の成果をもとに有意義な研究を展開していきたい。

Research Products

• [Journal Article] GATA transcription factors, SOX17 and TFAP2C, drive the human germ-cell specification program (2021)
  • Author(s): Kojima Yoji、Yamashiro Chika、Murase Yusuke、Yabuta Yukihiro、Okamoto Ikuhiro、Iwatani Chizuru、Tsuchiya Hideaki、Nakaya Masataka、Tsukiyama Tomoyuki、Nakamura Tomonori、Yamamoto Takuya、Saitou Mitinori
  • Journal Title: Life Science Alliance Volume: 4 Issue: 5 Pages: 202000974-202000974
  • DOI: 10.26508/lsa.202000974
  • NAID: 120006979739
  • Peer Reviewed / Open Access
• [Journal Article] Capturing human trophoblast development with naive pluripotent stem cells in vitro (2021)
  • Author(s): S Io M Kabata Y Lemura N Morone A Minagawa B Wong I Okamoto T Nakamura Y Kojima C Iwatani H Tsuchiya B Kawswandy E KondohS Kaneko K Woltjen M Saitou T Yamanoto M Mandai Y Takashima
  • Journal Title: Cell Stem Cell Volume: 21 Issue: 6 Pages: 10-10
  • DOI: 10.1016/j.stem.2021.03.013
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Cyclosporin A and FGF signaling support the proliferation/survival of mouse primordial germ cell-like cells in vitro† (2020)
  • Author(s): Ohta Hiroshi、Yabuta Yukihiro、Kurimoto Kazuki、Nakamura Tomonori、Murase Yusuke、Yamamoto Takuya、Saitou Mitinori
  • Journal Title: Biology of Reproduction Volume: 104 Issue: 2 Pages: 344-360
  • DOI: 10.1093/biolre/ioaa195
  • Peer Reviewed / Open Access
• [Journal Article] ZGLP1 is a determinant for the oogenic fate in mice (2020)
  • Author(s): Nagaoka So I.、Nakaki Fumio、Miyauchi Hidetaka、Nosaka Yoshiaki、Ohta Hiroshi、Yabuta Yukihiro、Kurimoto Kazuki、Hayashi Katsuhiko、Nakamura Tomonori、Yamamoto Takuya、Saitou Mitinori
  • Journal Title: Science Volume: 367 Issue: 6482 Pages: 4115-4115
  • DOI: 10.1126/science.aaw4115
  • NAID: 120006850276
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Long‐term expansion with germline potential of human primordial germ cell‐like cells in?vitro (2020)
  • Author(s): Murase Yusuke、Yabuta Yukihiro、Ohta Hiroshi、Yamashiro Chika、Nakamura Tomonori、Yamamoto Takuya、Saitou Mitinori
  • Journal Title: The EMBO Journal Volume: 39 Issue: 21 Pages: 104929-104929
  • DOI: 10.15252/embj.2020104929
  • Peer Reviewed
• [Journal Article] Induction of the germ cell fate from pluripotent stem cells in cynomolgus monkeys. (2020)
  • Author(s): Sakai Y, Nakamura T, Okamoto I, Gyobu-Motani S, Ohta H, Yabuta Y, Tsukiyama T, Iwatani C, Tsuchiya H, Ema M, Morizane A, Takahashi J, Yamamoto T, Saitou M.
  • Journal Title: Biol Reprod Volume: 102(3) Issue: 3 Pages: 620-638
  • DOI: 10.1093/biolre/ioz205
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Generation of human oogonia from induced pluripotent stem cells in vitro. (2018)
  • Author(s): 30.Yamashiro C, Sasaki K, Yabuta Y, Kojima Y, Nakamura T, Okamoto I, Yokobayashi S, Murase Y, Ishikura Y, Shirane K, Sasaki H, Yamamoto T, Saitou M.
  • Journal Title: Science Volume: 362(6412) Issue: 6412 Pages: 356-360
  • DOI: 10.1126/science.aat1674
  • Peer Reviewed
• [Presentation] 霊長類特異的多能性状態の解明 (2020)
  • Author(s): Tomonori Nakamura* , Yusuke Imoto, Yasuaki Hiraoka, Mitinori Saitou
  • Organizer: 第53回発生生物学会年会
  • Invited
• [Presentation] Identifying cells hidden by curse of dimensionality (2020)
  • Author(s): Tomonori Nakamura
  • Organizer: The 1st ASHBi SignAC international workshop 2021
  • Invited
• [Presentation] Dissecting the spectrum of pluripotency in primates (2020)
  • Author(s): Tomonori Nakamura
  • Organizer: 第５３回日本発生生物学会年会
  • Int'l Joint Research / Invited
• [Presentation] Induction of the Germ-Cell Fate from Pluripotent Stem Cells in Cynomolgus Monkeys (2019)
  • Author(s): Tomonori Nakamura
  • Organizer: 第４２回日本分子生物学会年会
  • Int'l Joint Research / Invited
• [Presentation] 次元の呪い~高次元観測データにおけるノイズとノイズ低減解析法~ (2019)
  • Author(s): 中村友紀
  • Organizer: 第６回六甲医学研究会
  • Invited
• [Patent(Industrial Property Rights)] 観測ノイズ削減法の開発 (2020)
  • Inventor(s): 井元祐介、中村友紀、平岡、斎藤通紀
  • Industrial Property Rights Holder: 井元祐介、中村友紀、平岡、斎藤通紀
  • Industrial Property Rights Type: 特許
  • Filing Date: 2020