Research on long-term storage of mammalian eggs and early embryos at room temperature ~for application to genetic resources preservation~

• Project/Area Number: 18K05691
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Review Section: Basic Section 39060:Conservation of biological resources-related
• Research Institution: Juntendo University
• Principal Investigator: TADA Norihiro 順天堂大学, 大学院医学研究科, 先任准教授 (50338315)
• Project Period (FY): 2018-04-01 – 2023-03-31
• Project Status: Completed (Fiscal Year 2022)
• Budget Amount: ¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000); Fiscal Year 2021: ¥910,000 (Direct Cost: ¥700,000、Indirect Cost: ¥210,000); Fiscal Year 2020: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000); 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: マウス / 初期胚 / 真空乾燥 / 常温保存 / トレハロース / CRISPR/Cas9 / 電気穿孔法 / バイオリソース / 卵子 / 室温保存 / エレクトロポレーション / 受精卵 / 遺伝資源保存

Outline of Final Research Achievements

We generated knock-in (KI) mice expressing a trehalose transporter that can take up trehalose, which has a strong biocomponent-protective effect on cells during desiccation, into cells by genome editing using CRISPR/Cas9 system. Using oocytes and early embryos from KI mice, we tried vacuum drying and room temperature storage under various conditions, but they did not survive after rehydration. In addition, we attempted to incorporate trehalose into the cells of early embryos by electroporation, but no protective effect was observed in vacuum-dried early embryos. Furthermore, after microinjection of trehalose into the cytoplasm of 1-cell stage eggs, they were allowed to develop into 2-cell stage embryos. No embryos were obtained.

Academic Significance and Societal Importance of the Research Achievements

卵子、初期胚のような生殖機能細胞はいずれも、真空乾燥・常温保存の試みは成されているが、今だ成功していない。精子では成功しているので、その違いがどこにあるのか、核のみならず細胞質及び細胞膜の乾燥に対する耐性機構が明らかになるので、学術的な意義は大きい。また現在、生殖機能細胞の保存はすべて凍結保存であるが、本研究で真空乾燥・常温保存の有効性を明らかにできれば、遺伝資源の開発・保全に極めて有用な手段となり得る。特に、液体窒素中での凍結保存については、液体窒素を補充しなければならず、危険物なので宅配便等での輸送が制限される。しかし、常温保存であれば郵便などで容易に送ることができる。

Research Products

• [Journal Article] Impaired GATE16-mediated exocytosis in exocrine tissues causes Sj?gren’s syndrome-like exocrinopathy (2022)
  • Author(s): Suzuki Akiko、Iwaya Chihiro、Ogata Kenichi、Yoshioka Hiroki、Shim Junbo、Tanida Isei、Komatsu Masaaki、Tada Norihiro、Iwata Junichi
  • Journal Title: Cellular and Molecular Life Sciences Volume: 79 Issue: 6 Pages: 307-307
  • DOI: 10.1007/s00018-022-04334-x
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Ptpn20 deletion in H-Tx rats enhances phosphorylation of the NKCC1 cotransporter in the choroid plexus: an evidence of genetic risk for hydrocephalus in an experimental study (2022)
  • Author(s): Xu Hanbing、Miyajima Masakazu、Nakajima Madoka、Ogino Ikuko、Kawamura Kaito、Akiba Chihiro、Kamohara Chihiro、Sakamoto Koichiro、Karagiozov Kostadin、Nakamura Eri、Tada Nobuhiro、Arai Hajime、Kondo Akihide
  • Journal Title: Fluids and Barriers of the CNS Volume: 19 Issue: 1 Pages: 1-15
  • DOI: 10.1186/s12987-022-00341-z
  • Peer Reviewed / Open Access
• [Journal Article] Impaired mitochondrial accumulation and Lewy pathology in neuron-specific FBXO7-deficient mice (2022)
  • Author(s): Noda Sachiko、Sato Shigeto、Fukuda Takahiro、Ueno Shinichi、Tada Norihiro、Hattori Nobutaka
  • Journal Title: Molecular Brain Volume: 15 Issue: 1 Pages: 54-54
  • DOI: 10.1186/s13041-022-00936-5
  • Peer Reviewed
• [Journal Article] Differentiation of enteric neural crest cells transplanted from SOX10-Venus mouse embryonic stem cells into the gut of the endothelin receptor B null mouse model (2022)
  • Author(s): Fujiwara N, Miyahara K, Nakazawa-Tanaka N, Oishi Y, Akazawa C, Tada N, Yamataka A
  • Journal Title: Pediatr Surg Int Volume: 39 Issue: 1 Pages: 18-18
  • DOI: 10.1007/s00383-022-05318-5
  • Peer Reviewed / Open Access
• [Journal Article] Interleukin-11-expressing fibroblasts have a unique gene signature correlated with poor prognosis of colorectal cancer. (2021)
  • Author(s): Nishina T, Deguchi Y, Ohshima D, Takeda W, Ohtsuka M, Shichino S, Ueha S, Yamazaki S, Kawauchi M, Nakamura E, Nishiyama C, Kojima Y, Adachi-Akahane S, Hasegawa M, Nakayama M, Oshima M, Yagita H, Shibuya K, Mikami T, Inohara N, Matsushima K, Tada N, Nakano H.
  • Journal Title: Nat Commun. Volume: 12 Issue: 1 Pages: 2281-2281
  • DOI: 10.1038/s41467-021-22450-3
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] FADS2 ablation exacerbates hepatic triacylglycerol and cholesterol accumulation in polyunsaturated fatty acid-depleted mice (2021)
  • Author(s): Y. Hayashi, HC. Lee-Okada, E. Nakamura, N. Tada, T. Yokomizo, Y. Fujiwara, I. Ichi
  • Journal Title: FEBS letters Volume: Accepted Issue: 14 Pages: 1920-1932
  • DOI: 10.1002/1873-3468.14134
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Homeostatic p62 levels and inclusion body formation in CHCHD2 knockout mice (2021)
  • Author(s): Sato Shigeto、Noda Sachiko、Torii Satoru、Amo Taku、Ikeda Aya、Funayama Manabu、Yamaguchi Junji、Fukuda Takahiro、Kondo Hiromi、Tada Norihiro、Arakawa Satoko、Watanabe Masahiko、Uchiyama Yasuo、Shimizu Shigeomi、Hattori Nobutaka
  • Journal Title: Human Molecular Genetics Volume: 30 Issue: 6 Pages: 443-453
  • DOI: 10.1093/hmg/ddab057
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Ependymal ciliary motion and their role in congenital hydrocephalus. (2021)
  • Author(s): K. Sakamoto, M. Nakajima, K. Kawamura, E. Nakamura, N. Tada, A. Kondo, H. Arai, M. Miyajima
  • Journal Title: Child's Nervous System Volume: 37(11) Issue: 11 Pages: 3355-3364
  • DOI: 10.1007/s00381-021-05194-9
  • Peer Reviewed / Open Access
• [Journal Article] Loss of Parkin contributes to mitochondrial turnover and dopaminergic neuronal loss in aged mice. (2020)
  • Author(s): Sachiko Noda; Takahiro Fukuda; Norihiro Tada; Yasuo Uchiyama; Keiji Tanaka; Nobutaka Hattori.
  • Journal Title: Neurobiology of Disease. Volume: 136 Pages: 104717-104717
  • Peer Reviewed
• [Journal Article] Aging-related motor function and dopaminergic neuronal loss in C57BL/6 mice. (2020)
  • Author(s): Sachiko Noda; Shigeto Sato; Takahiro Fukuda; Norihiro Tada; Nobutaka Hattori.
  • Journal Title: Molecular Brain Volume: 13 Pages: 46-46
  • Peer Reviewed
• [Journal Article] Low bone mineral density due to secondary hyperparathyroidism in the GlatmTg(CAG-A4GALT) mouse model of Fabry disease. (2020)
  • Author(s): H Maruyama, A Taguchi, M Mikane, H Lu, N Tada, M Ishijima, H Kaneko, M Kawai, S Goto, A Saito, R Ohashi, Y Nishikawa, S Ishii.
  • Journal Title: FASEB BioAdv. Volume: 2 Pages: 365-381
  • Peer Reviewed / Open Access
• [Journal Article] Characterization of starvation-induced autophagy in cerebellar Purkinje cells of pHluorin-mKate2-human LC3B transgenic mice. (2020)
  • Author(s): Juan Alejandro Oliva Trejo, Isei Tanida, Chigure Suzuki, Soichiro Kakuta, Norihiro Tada, and Yasuo Uchiyama.
  • Journal Title: Scientific Reports Volume: 10(1) Issue: 1 Pages: 9643-9643
  • DOI: 10.1038/s41598-020-66370-6
  • Peer Reviewed
• [Journal Article] Expression of leukotriene B4 receptor 1 defines functionally distinct DCs that control allergic skin inflammation. (2020)
  • Author(s): Tomoaki Koga, Fumiyuki Sasaki, Kazuko Saeki, Soken Tsuchiya, Toshiaki Okuno, Mai Ohba, Takako Ichiki, Satoshi Iwamoto, Hirotsugu Uzawa, Keiko Kitajima, Chikara Meno, Eri Nakamura, Norihiro Tada, Yoshinori Fukui, Junichi Kikuta, Masaru Ishii, Yukihiko Sugimoto, and Mitsuyoshi Nakao, Takehiko Yokomizo.
  • Journal Title: Cellular & Molecular Immunology Volume: 8 Issue: 6 Pages: 1-13
  • DOI: 10.1038/s41423-020-00559-7
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Periostin deletion suppresses late-phase response in mouse experimental allergic conjunctivitis (2019)
  • Author(s): Asada Y, Okano M, Ishida W, Iwamoto S, Fukuda K, Hirakata T, Tada N, Fukushima A, Ebihara N, Kudo A, Matsuda A.
  • Journal Title: Allergology International Volume: 68 Issue: 2 Pages: 233-239
  • DOI: 10.1016/j.alit.2018.09.007
  • NAID: 130007633580
  • ISSN: 1323-8930, 1440-1592
  • Peer Reviewed / Open Access
• [Journal Article] Compromised vitality of spermatozoa after contact with colonic mucosa in mice: implications for fertility in colon vaginoplasty patients. (2019)
  • Author(s): Kosaka S, Takeda M, Ochi T, Miyahara K, Nakamura E, Tada N, Lane GJ, Yamataka A.
  • Journal Title: Pediatric Surgery International Volume: 35(1) Issue: 1 Pages: 71-75
  • DOI: 10.1007/s00383-018-4377-3
  • Peer Reviewed
• [Journal Article] Induced Pluripotent Stem Cells Reprogrammed with Three Inhibitors Show Accelerated Differentiation Potentials with High Levels of 2-Cell Stage Marker Expression (2019)
  • Author(s): Nishihara Koji、Shiga Takahiro、Nakamura Eri、Akiyama Tomohiko、Sasaki Takashi、Suzuki Sadafumi、Ko Minoru S.H.、Tada Norihiro、Okano Hideyuki、Akamatsu Wado
  • Journal Title: Stem Cell Reports Volume: 12 Issue: 2 Pages: 305-318
  • DOI: 10.1016/j.stemcr.2018.12.018
  • Peer Reviewed / Open Access
• [Journal Article] Hlf marks the developmental pathway for hematopoietic stem cells but not for erythro-myeloid progenitors., (2019)
  • Author(s): Tomomasa Yokomizo, Naoki Watanabe, Terumasa Umemoto, Junichi Matsuo, Ryota Harai, Yoshihiko Kihara, Eri Nakamura, Norihiro Tada, Tomohiko Sato, Tomoiku Takaku, Akihiko Shimono, Hitoshi Takizawa, Naomi Nakagata, Seiichi Mori, Mineo Kurokawa, Daniel Tenen, Motomi Osato, Toshio Suda, and Norio Komatsu.
  • Journal Title: J Exp Med Volume: 216 Issue: 7 Pages: 1599-1614
  • DOI: 10.1084/jem.20181399
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] KIF11 as a Potential Marker of Spermatogenesis Within Mouse Seminiferous Tubule Cross-sections (2019)
  • Author(s): Miki Hara-Yokoyama, Hidetake Kurihara, Shozo Ichinose, Hironori Matsuda, Shizuko Ichinose, Masaru Kurosawa, Norihiro Tada, Chihiro Iwahara, Kazue Terasawa, Katarzyna A Podyma-Inoue, Koichi Furukawa , Kazuhisa Iwabuchi
  • Journal Title: Journal of Histochemistry & Cytochemistry Volume: 67 Issue: 11 Pages: 813-824
  • DOI: 10.1369/0022155419871027
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] FcgRIIB on B cells and myeloid cells modulates B cell activation and autoantibody responses via different but synergistic pathways in Lupus-Prone Yaa mice. (2018)
  • Author(s): Qingshum Lin, Mareki Ohtsuji, Hirofumi Amano, Hiromichi Tsurui, Norihiro Tada, Ryota Sato, Hidehiro Fukuyama, Hiroyuki Nishimura, J Sjef Verbeek, Sachiko Hisose
  • Journal Title: Journal of Immunology Volume: 201 Issue: 11 Pages: 3199-3210
  • DOI: 10.4049/jimmunol.1701487
  • Peer Reviewed
• [Presentation] 精子における脂肪酸輸送体 (FATP1)の役割の解明 (2022)
  • Author(s): 陣内 ひろみ、佐伯 和子、中村 衣里、多田 昇弘、李 賢哲、横溝 岳彦
  • Organizer: 第64回日本脂質生化学会
• [Presentation] 雄性生殖における脂肪酸輸送体(FATP1)の役割の解明 (2022)
  • Author(s): 陣内 ひろみ, 佐伯 和子,中村 衣里, 多田 昇弘, 李 賢哲, 横溝 岳彦
  • Organizer: 第95回日本生化学会
• [Presentation] 複数のRyR2変異マウスモデル系統を用いたカテコラミン誘発性多型性心室頻拍（CPVT）に対する抗不整脈薬の評価 (2022)
  • Author(s): 呉林 なごみ，児玉 昌美，村山 尚，杉原 匡美，石井 光一郎，岡部 雄太，村越 伸行，三浦 綾，西尾 元，井上-上野 由紀子，井上 高良，野口 悟，中村 衣里，金井 富士夫，多田 昇弘，櫻井 隆
  • Organizer: 第96回日本薬理学会年会
• [Presentation] C57BL/6マウスにおける加齢に伴う運動機能とドーパミン作動性ニューロンの変性 (2021)
  • Author(s): 野田幸子、佐藤栄人、福田隆浩、多田昇弘、服部信孝
  • Organizer: 第68回日本実験動物学会総会（Web開催）
• [Presentation] 水頭症を発症する微小管モーター蛋白質ダイニンDnah5欠損マウスの病態解析 (2021)
  • Author(s): 中村衣里、荻野郁子、川村海渡、阪本浩一朗、中島円、宮原克、多田昇弘、宮嶋雅一
  • Organizer: 第68回日本実験動物学会総会（Web開催）
• [Presentation] CHCHD2欠損パーキンソン病モデルマウスの解析 (2021)
  • Author(s): 佐藤栄人、野田幸子、多田昇弘、服部信孝
  • Organizer: 第68回日本実験動物学会総会（Web開催）
• [Presentation] 微小管モータータンパク質ダイニンDnah5の欠損マウスは繊毛形成不全を起こし水頭症を発症する (2020)
  • Author(s): 中村衣里、荻野郁子、川村海渡、中島円、宮原克、吉田光孝、多田昇弘、宮嶋雅一
  • Organizer: 第67回日本実験動物学会総会
• [Presentation] 加齢マウスにみられる運動機能とドーパミン神経細胞の変性について (2019)
  • Author(s): 野田幸子、佐藤栄人、多田昇弘、福田隆浩、服部信孝
  • Organizer: 第66回日本実験動物学会総会
• [Presentation] ゲノム編集技術を用いた水頭症モデルマウスの作製 (2019)
  • Author(s): 中村衣里、荻野郁子、川村海渡、中島円、宮原克、多田昇弘、宮嶋雅一
  • Organizer: 第66回日本実験動物学会総会
• [Presentation] 新動物実験施設への移転に向けた取り組み (2019)
  • Author(s): 山口裕之、多田昇弘
  • Organizer: 第53回日本実験動物技術者協会総会
• [Presentation] マウスにおけるピエゾマイクロインジェクション法を用いた効率的な個体復元の試み (2018)
  • Author(s): 中村衣里、多田昇弘
  • Organizer: 第65回日本実験動物学会総会
• [Presentation] Decreased viability/motility of spermatozoa after contact with colonic mucosa in mice may be implicated in infertility of cloacal malformation patients after colon vaginoplasty (2018)
  • Author(s): Kosaka S, Takeda M, Miyahara K, Nakamura E, Norihiro Tada N, Lane G, Yamataka A
  • Organizer: The Pacific Association of Pediatric Surgeons
  • Int'l Joint Research
• [Presentation] Compromised vitality of spermatozoa after contact with colonic mucosa in mice. Implications for fertility in colon vaginoplasty patients. (2018)
  • Author(s): Seitaro Kosaka, Masahiro Takeda, Ochi Takanori, Katsumi Miyahara, Eri Nakamura, Norihiro Tada, Geoffrey Lane, Atsuyuki Yamataka
  • Organizer: 31st INTERNATIONAL SYMPOSIUM
  • Int'l Joint Research
• [Book] ゲノム編集技術を応用した製品開発とその実用化 (2021)
  • Author(s): 多田昇弘 他 共著
  • Total Pages: 602
  • Publisher: （株）技術情報協会
• [Patent(Industrial Property Rights)] パーキン病モデル非ヒト動物 (2019)
  • Inventor(s): 佐藤栄人、野田幸子、多田昇弘、服部信孝
  • Industrial Property Rights Holder: 佐藤栄人、野田幸子、多田昇弘、服部信孝
  • Industrial Property Rights Type: 特許
  • Industrial Property Number: 2019-154898
  • Filing Date: 2019
  • Acquisition Date: 2019