Reconstituion in vitro of primate germ cell development

• Project/Area Number: 21K15112
• Research Category: Grant-in-Aid for Early-Career Scientists
• Allocation Type: Multi-year Fund
• Review Section: Basic Section 44020:Developmental biology-related
• Research Institution: Institute of Physical and Chemical Research (2022); Kyushu University (2021)
• Principal Investigator: Yoshimatsu Sho 国立研究開発法人理化学研究所, 脳神経科学研究センター, 客員研究員 (00882713)
• Project Period (FY): 2021-04-01 – 2023-03-31
• Project Status: Completed (Fiscal Year 2022)
• Budget Amount: ¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000); Fiscal Year 2022: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000); Fiscal Year 2021: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
• Keywords: 生殖細胞 / マーモセット / 霊長類 / シングルセル / 多能性幹細胞 / 卵子 / ゲノム編集 / レポーター / 再構築

Outline of Research at the Start

生殖細胞系譜は次世代に遺伝情報を伝達し、その系譜分化の機構究明は進化生物学・発生学上の重要課題とされてきた。近年、マウス多能性幹細胞から生殖系列細胞を誘導し産仔形成能をもつ機能的配偶子が作出されたという成果は、「多能性幹細胞からのin vitro配偶子形成」という革新的生殖技術に繋がり、ヒト細胞についてもin vitro配偶子形成研究の新領域を拓いた。しかし生殖医療への応用を踏まえた場合、実験的検証を可能とする非ヒト霊長類を用いた培養系の開発が求められる。そこで本研究では小型霊長類のマーモセットを対象として、多能性幹細胞から配偶子分化をin vitroで再現する培養系の確立を最終目標とする。

Outline of Final Research Achievements

In this study, single cell analysis of germline cells from marmosets allowed us to identify primate-specific gene expression profiles, a major step toward future reconstruction in culture. Also, we generated reporter embryonic stem cell lines of marmosets harboring gonadal somatic cell-reporters. It would promote further studies for in vitro reconstitution of functional gametes from marmosets.

Academic Significance and Societal Importance of the Research Achievements

本研究の継続よって、生殖細胞の再構築を検討する上で、近年深刻になっている少子高齢化や、不妊治療に対する根本的な解決策を創出できることがきたいされる。

Research Products

• [Int'l Joint Research] シンシナティ小児病院医療センター(米国)
• [Int'l Joint Research] German Primate Center(ドイツ)
• [Int'l Joint Research] Cincinnati Children's Hospital(米国)
• [Journal Article] Highly efficient induction of functionally mature excitatory neurons from feeder-free human ES/iPS cells (2023)
  • Author(s): Zhou Zhi、Kakegawa Wataru、Fujimori Koki、Sho Misato、Shimamura Rieko、Supakul Sopak、Yoshimatsu Sho、Kohyama Jun、Yuzaki Michisuke、Okano Hideyuki
  • Journal Title: bioRxiv Volume: N/A
  • DOI: 10.1101/2023.02.10.528087
  • Open Access
• [Journal Article] Generation of a Tyrosine Hydroxylase (TH)-2A-Cre Knock-In Non-Human Primate Model by Homology Directed Repair-Biased CRISPR-Cas9 Genome Editing (2023)
  • Author(s): Yoshimatsu Sho、Okahara Junko、Yohie Junko、Igarashi Yoko、Nakajima Ryusuke、Sanosaka Tsukasa、Qian Emi、Kobayashi Hiroya、Morimoto Satoru、Kishi Noriyuki、Pillis Devin M.、Malik Punam、Noce Toshiaki、Okano Hideyuki
  • Journal Title: SSRN Volume: N/A
  • DOI: 10.2139/ssrn.4319536
  • Open Access
• [Journal Article] Early development of the cochlea of the common marmoset, a non-human primate model (2022)
  • Author(s): Hosoya Makoto、Fujioka Masato、Okahara Junko、Yoshimatsu Sho、Okano Hideyuki、Ozawa Hiroyuki
  • Journal Title: Neural Development Volume: 17 Issue: 1 Pages: 6-6
  • DOI: 10.1186/s13064-022-00162-8
  • Peer Reviewed / Open Access
• [Journal Article] Critical roles of FGF, RA, and WNT signaling in the development of human otic placode and subsequent lineages in a dish (2022)
  • Author(s): Saeki, T., Yoshimatsu, S., Ishikawa, M., Shin, JW., Hon, C-C., Shibata, S., Hosoya, M., Saegusa, C., Ogawa, K., Fujioka, M., Okano, H.
  • Journal Title: Regen Ther Volume: 20 Pages: 165-186
  • DOI: 10.1016/j.reth.2022.04.008
  • Peer Reviewed / Open Access
• [Journal Article] A New Horizon in Reproductive Research with Pluripotent Stem Cells: Successful In Vitro Gametogenesis in Rodents, Its Application to Large Animals, and Future In Vitro Reconstitution of Reproductive Organs Such as “Uteroid” and “Oviductoid” (2022)
  • Author(s): Yoshimatsu Sho、Kisu Iori、Qian Emi、Noce Toshiaki
  • Journal Title: Biology Volume: 11 Issue: 7 Pages: 987-987
  • DOI: 10.3390/biology11070987
  • Peer Reviewed / Open Access
• [Journal Article] Single transcription factor leads pluripotent stem cells to functional microglia (2022)
  • Author(s): Sonn Iki, Ozaki Fumiko, Yoshimatsu Sho, Morimoto Satoru, Watanabe Hirotaka, Okano Hideyuki
  • Journal Title: Inflammation and Regeneration Volume: (in press) Issue: 1 Pages: 1-1
  • DOI: 10.1186/s41232-022-00201-1
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Step‐by‐step protocols for non‐viral derivation of transgene‐free induced pluripotent stem cells from somatic fibroblasts of multiple mammalian species (2022)
  • Author(s): Yoshimatsu Sho、Yamazaki Atsushi、Edamura Kazuya、Koushige Yuko、Shibuya Hisashi、Qian Emi、Sato Tsukika、Okahara Junko、Kishi Noriyuki、Noce Toshiaki、Yamaguchi Yoshifumi、Okano Hideyuki
  • Journal Title: Development, Growth & Differentiation Volume: 64 Issue: 6 Pages: 325-341
  • DOI: 10.1111/dgd.12798
  • Peer Reviewed
• [Journal Article] Multimodal analyses of a non-human primate model harboring mutant amyloid precursor protein transgenes driven by the human EF1α promoter. (2022)
  • Author(s): Yoshimatsu S, Seki F, Okahara J, Watanabe H, Sasaguri H, Haga Y, Hata JI, Sanosaka T, Inoue T, Mineshige T, Lee CY, Shinohara H, Kurotaki Y, Komaki Y, Kishi N, Murayama AY, Nagai Y, Minamimoto T, Yamamoto M, Nakajima M, Zhou Z, Nemoto A, Sato T, Ikeuchi T, Sahara N, Morimoto S, Shiozawa S, Saido TC, Sasaki E, Okano H
  • Journal Title: Neuroscience Research Volume: 185 Pages: 49-61
  • DOI: 10.1016/j.neures.2022.08.008
  • Peer Reviewed / Open Access
• [Journal Article] Attempts for deriving extended pluripotent stem cells from common marmoset embryonic stem cells (2022)
  • Author(s): Yoshimatsu S, Nakajima M, Sonn I, Natsume R, Sakimura K, Nakatsukasa E, Sasaoka T, Nakamura M, Serizawa T, Sato T, Sasaki E, Deng H, Okano H
  • Journal Title: Genes to Cells Volume: 28(2) Issue: 2 Pages: 156-169
  • DOI: 10.1111/gtc.13000
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Homologous Recombination-Enhancing Factors Identified by Comparative Transcriptomic Analyses of Pluripotent Stem Cell of Human and Common Marmoset (2022)
  • Author(s): Yoshimatsu Sho、Nakajima Mayutaka、Qian Emi、Sanosaka Tsukasa、Sato Tsukika、Okano Hideyuki
  • Journal Title: Cells Volume: 11 Issue: 3 Pages: 360-360
  • DOI: 10.3390/cells11030360
  • Peer Reviewed / Open Access
• [Journal Article] Early development of the cochlea of the common marmoset, a non-human primate model (2022)
  • Author(s): Hosoya Makoto, Fujioka Msato, Okahara Junko, Yoshimatsu Sho, Okano Hideyuki, Ozawa Hiroyuki
  • Journal Title: Neural Development Volume: (in press)
  • Peer Reviewed / Open Access
• [Journal Article] Critical roles of FGF, RA, and WNT signaling in the development of human otic placode and subsequent lineages in a dish (2022)
  • Author(s): Saeki Tsubasa, Yoshimatsu Sho, Hosoya Makoto, Saegusa Chika, Ogawa Kaoru, Fujioka Masato, Okano Hideyuki
  • Journal Title: Regenerative Therapy Volume: (in press)
  • Peer Reviewed / Open Access
• [Journal Article] Accelerated neuronal ageing in vitro ~Melting Watch~ (2022)
  • Author(s): Inagaki Emi, Imaizumi Kent, Yoshimatsu Sho, Okano Hideyuki
  • Journal Title: Frontiers in Neuroscience Volume: (in press)
  • Peer Reviewed / Open Access
• [Journal Article] Generation of a control human induced pluripotent stem cell line using the defective and persistent Sendai virus vector system (2021)
  • Author(s): Zhou Zhi、Yoshimatsu Sho、Qian Emi、Ishikawa Mitsuru、Sato Tsukika、Ohtaka Manami、Nakanishi Mahito、Okano Hideyuki
  • Journal Title: Stem Cell Research Volume: 56 Pages: 102549-102549
  • DOI: 10.1016/j.scr.2021.102549
  • Peer Reviewed / Open Access
• [Journal Article] Establishment of an induced pluripotent stem cell line from a female domestic ferret (Mustela putorius furo) with an X chromosome instability (2021)
  • Author(s): Yoshimatsu Sho、Murakami Rei、Nakajima Mayutaka、Sato Tsukika、Kawasaki Hiroshi、Okano Hideyuki
  • Journal Title: Stem Cell Research Volume: 53 Pages: 102385-102385
  • DOI: 10.1016/j.scr.2021.102385
  • Peer Reviewed / Open Access
• [Journal Article] Establishing an induced pluripotent stem cell line from neonatal common marmoset fibroblasts by an all-in-one episomal vector approach (2021)
  • Author(s): Yoshimatsu Sho、Qian Emi、Sato Tsukika、Yamamoto Masafumi、Ishikawa Mitsuru、Okano Hideyuki
  • Journal Title: Stem Cell Research Volume: 53 Pages: 102380-102380
  • DOI: 10.1016/j.scr.2021.102380
  • Peer Reviewed / Open Access
• [Journal Article] Non-viral derivation of a transgene-free induced pluripotent stem cell line from a male beagle dog (2021)
  • Author(s): Yoshimatsu Sho、Edamura Kazuya、Yoshii Yumi、Iguchi Aozora、Kondo Hirotaka、Shibuya Hisashi、Sato Tsukika、Shiozawa Seiji、Okano Hideyuki
  • Journal Title: Stem Cell Research Volume: 53 Pages: 102375-102375
  • DOI: 10.1016/j.scr.2021.102375
  • Peer Reviewed / Open Access
• [Journal Article] Generation of a common marmoset embryonic stem cell line CMES40-OC harboring a POU5F1 (OCT4)-2A-mCerulean3 knock-in reporter allele (2021)
  • Author(s): Yoshimatsu S, Murakami R, Sato T, Saeki T, Yamamoto M, Sasaki E, Noce T, Okano H
  • Journal Title: Stem Cell Research Volume: 53 Pages: 102308-102308
  • DOI: 10.1016/j.scr.2021.102308
  • Peer Reviewed / Open Access
• [Journal Article] Non-viral Induction of Transgene-free iPSCs from Somatic Fibroblasts of Multiple Mammalian Species. (2021)
  • Author(s): Yoshimatsu S, Nakajima M, Iguchi A, Sanosaka T, Sato T, Nakamura M, Nakajima R, Arai E, Ishikawa M, Imaizumi K, Watanabe H, Okahara J, Noce T, Takeda Y, Sasaki E, Behr R, Edamura K, Shiozawa S, Okano H.
  • Journal Title: Stem Cell Reports Volume: 16 Issue: 4 Pages: 754-770
  • DOI: 10.1016/j.stemcr.2021.03.002
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] クローズアップ実験法series337「さまざまな動物種から高品質なiPS細胞を作出する方法」 (2021)
  • Author(s): 吉松祥, 岡野栄之
  • Journal Title: 実験医学 Volume: 39
• [Presentation] A new horizon of marmoset research using highly efficient knock-in and pluripotent stem cell-based technologies (2021)
  • Author(s): 1)Yoshimatsu Sho, Okahara Junko, Nemoto Akisa, Sato Tsukika, Kishi Noriuki, Noce Toshiaki, Hayashi Katsuhiko, Okano Hideyuki
  • Organizer: ISSCR/JSMR International Symposium 2021
  • Int'l Joint Research / Invited
• [Presentation] 高効率ノックイン技術を用いたマーモセット研究の新しい展開 (2021)
  • Author(s): 吉松祥, 岡原純子, 根本晶沙, 佐藤月花, 岸憲幸, 野瀬俊明, 岡野栄之
  • Organizer: 第44回日本神経科学大会 / CJK第1回国際会議
• [Funded Workshop] ISSCR/JSMR International Symposium 2021 (2021)