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The attempt to improve the developmental competence of oocytes using in vitro growth model of bovine follicles and the application for the production of wildlife oocytes

Research Project

Project/Area Number 16K08043
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeMulti-year Fund
Section一般
Research Field Veterinary medical science
Research InstitutionHokkaido University

Principal Investigator

Nagano Masashi  北海道大学, 獣医学研究院, 准教授 (70312402)

Research Collaborator YANAGAWA Yojiro  
SAKAGUCHI Kenichiro  
KANNO Chihiro  
Project Period (FY) 2016-04-01 – 2019-03-31
Project Status Completed (Fiscal Year 2018)
Budget Amount *help
¥4,940,000 (Direct Cost: ¥3,800,000、Indirect Cost: ¥1,140,000)
Fiscal Year 2018: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000)
Fiscal Year 2017: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000)
Fiscal Year 2016: ¥2,470,000 (Direct Cost: ¥1,900,000、Indirect Cost: ¥570,000)
Keywords卵子 / 体外受精 / 体外発育培養 / 細胞質成熟 / 卵胞 / 体外成熟培養 / 卵胞発育
Outline of Final Research Achievements

To clarify the relationship between the steroidogenesis and the acquisition of developmental competence of bovine oocytes during follicular development or degradation, I cultured the developing bovine oocytes surrounded by granulosa cell layers. When FSH and BMP-4 were added to culture media of oocytes, luteinization of granulosa cells were inhibited but the developmental competence of oocytes decreased. Then I try to develop the new co-culture system with oocyte-granulosa cell complex (OGC) and theca cells which secrets several growth factors having the function for the development and the acquisition of developmental competence of oocytes. When OGC cultured with theca cells, theca and granulosa cells secreted androstenedione and estradiol-17β, respectively. And oocytes acquired the developmental competence to transferrable embryos. However, the developmental rate was low. We should develop the culture system which can support the function of theca and granulosa cells.

Academic Significance and Societal Importance of the Research Achievements

体外受精は家畜生産およびヒトの不妊治療分野で広く用いられているが作製された胚の発生能は低い。本研究の成果は卵子が発生能を獲得する過程を体外で明らかにする培養系の開発につながり、発生能の高い胚の体外生産にも資する。すなわち、家畜生産効率や不妊治療成功率の向上につながり、絶滅の危機にある野生動物の増殖にも寄与できる。

Report

(4 results)
  • 2018 Annual Research Report   Final Research Report ( PDF )
  • 2017 Research-status Report
  • 2016 Research-status Report
  • Research Products

    (35 results)

All 2019 2018 2017 2016

All Journal Article (10 results) (of which Int'l Joint Research: 4 results,  Peer Reviewed: 10 results,  Open Access: 6 results,  Acknowledgement Compliant: 2 results) Presentation (25 results) (of which Int'l Joint Research: 4 results,  Invited: 1 results)

  • [Journal Article] Astaxanthin improves the developmental competence of in vitro-grown oocytes and modifies the steroidogenesis of granulosa cells derived from bovine early antral follicles2019

    • Author(s)
      Abdel-Ghani M. A.、Yanagawa Y.、Balboula A. Z.、Sakaguchi K.、Kanno C.、Katagiri S.、Takahashi M.、Nagano M.
    • Journal Title

      Reproduction, Fertility and Development

      Volume: 31 Issue: 2 Pages: 272-272

    • DOI

      10.1071/rd17527

    • Related Report
      2018 Annual Research Report
    • Peer Reviewed / Int'l Joint Research
  • [Journal Article] Effects of pre-maturational culture duration on developmental competence of bovine small-sized oocytes2018

    • Author(s)
      ABDEL-GHANI Mohammed. A.、SAKAGUCHI Kenichiro、KANNO Chihiro、YANAGAWA Yojiro、KATAGIRI Seiji、NAGANO Masashi
    • Journal Title

      Journal of Reproduction and Development

      Volume: 64 Issue: 4 Pages: 365-369

    • DOI

      10.1262/jrd.2018-004

    • NAID

      130007437311

    • ISSN
      0916-8818, 1348-4400
    • Related Report
      2018 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Effect of a single epidural administration of follicle-stimulating hormone via caudal vertebrae on superstimulation for <i>in vivo</i> and <i>in vitro</i> embryo production in Japanese black cows2018

    • Author(s)
      SAKAGUCHI Kenichiro、IDETA Atsushi、YANAGAWA Yojiro、NAGANO Masashi、KATAGIRI Seiji、KONISHI Masato
    • Journal Title

      Journal of Reproduction and Development

      Volume: 64 Issue: 5 Pages: 451-455

    • DOI

      10.1262/jrd.2018-007

    • NAID

      130007496986

    • ISSN
      0916-8818, 1348-4400
    • Related Report
      2018 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Effect of astaxanthin addition to an individual culture system for in vitro maturation of bovine oocytes on accumulation of reactive oxygen species and mitochondrial activity2018

    • Author(s)
      Erdenetogtokh, P., Kanno, C., Sakaguchi, K., Yanagawa, Y., Katagiri, S. and Nagano, M
    • Journal Title

      Japanese Journal of Veterinary Research

      Volume: 66 Pages: 325-329

    • NAID

      120006539270

    • Related Report
      2018 Annual Research Report
    • Peer Reviewed / Open Access
  • [Journal Article] Relationship between the antral follicle count in bovine ovaries from a local abattoir and steroidogenesis of granulosa cells cultured as oocyte-cumulus-granulosa complexes2018

    • Author(s)
      SAKAGUCHI Kenichiro、TANIDA Takashi、ABDEL-GHANI Mohammed A.、KANNO Chihiro、YANAGAWA Yojiro、KATAGIRI Seiji、NAGANO Masashi
    • Journal Title

      Journal of Reproduction and Development

      Volume: 64 Issue: 6 Pages: 503-510

    • DOI

      10.1262/jrd.2018-080

    • NAID

      130007530498

    • ISSN
      0916-8818, 1348-4400
    • Related Report
      2018 Annual Research Report
    • Peer Reviewed / Open Access / Int'l Joint Research
  • [Journal Article] Relationship between in vitro growth of bovine oocytes and steroidogenesis of granulosa cells cultured in medium supplemented with bone morphogenetic protein-4 and follicle stimulating hormone2017

    • Author(s)
      Sakaguchi, K., Huang, W., Yang, Y., Yanagawa, Y., Nagano, M.*
    • Journal Title

      Theriogenology

      Volume: 印刷中 Pages: 113-123

    • DOI

      10.1016/j.theriogenology.2017.04.030

    • NAID

      120006459312

    • Related Report
      2017 Research-status Report 2016 Research-status Report
    • Peer Reviewed / Int'l Joint Research / Acknowledgement Compliant
  • [Journal Article] Extension of the culture period for the in vitro growth of bovine oocytes in the presence of bone morphogenetic protein-4 increases oocyte diameter, but impairs subsequent developmental competence2017

    • Author(s)
      Yang, Y., Kanno, C., Sakaguchi, K., Yanagawa, Y., Katagiri, S. and Nagano, M.
    • Journal Title

      Animal Science Journal

      Volume: 88 Issue: 11 Pages: 1686-1691

    • DOI

      10.1111/asj.12841

    • NAID

      120006539247

    • Related Report
      2017 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] Comparison of sperm subpopulation structures in first and second ejaculated semen from Japanese black bulls by a cluster analysis of sperm motility evaluated by a CASA system2017

    • Author(s)
      Kanno, C., Sakamoto, K. Q., Yanagawa, Y., Takahashi, Y., Katagiri, S. and Nagano, M.
    • Journal Title

      Journal of Veterinary Medical Science

      Volume: 79 Issue: 8 Pages: 1359-1365

    • DOI

      10.1292/jvms.17-0012

    • NAID

      130005901355

    • ISSN
      0916-7250, 1347-7439
    • Related Report
      2017 Research-status Report
    • Peer Reviewed / Open Access
  • [Journal Article] Effects of semen preservation procedure in egg yolk-tris based extender on bull spermatozoa characteristics2017

    • Author(s)
      Kanno, C., Yanagawa, Y., Takahashi, Y., Katagiri, S. and Nagano, M.
    • Journal Title

      Japanese Journal of Veterinary Research

      Volume: 65 Pages: 151-157

    • NAID

      120006346123

    • Related Report
      2017 Research-status Report
    • Peer Reviewed
  • [Journal Article] Extension of the culture period for the in vitro growth of bovine oocytes in the presence of bone morphogenetic protein-4 increases oocyte diameter, but impairs subsequent developmental competence2017

    • Author(s)
      Yang, Y., Kanno, C., Sakaguchi, K., Yanagawa, Y., Katagiri, S., Nagano, M.*
    • Journal Title

      Animal Science Journal

      Volume: 印刷中

    • NAID

      120006539247

    • Related Report
      2016 Research-status Report
    • Peer Reviewed / Acknowledgement Compliant
  • [Presentation] Effects of antral follicle count in ovaries on follicular development and endocrine dynamics of FSH and steroid hormones in cattle2019

    • Author(s)
      Sakaguchi K, Yanagawa Y, Yoshioka K, Suda Tomoko, Kawano Kohei, Katagiri S, Masashi N
    • Organizer
      45th Annual Conference of the International Embryo Technology Society
    • Related Report
      2018 Annual Research Report
    • Int'l Joint Research
  • [Presentation] 卵胞刺激ホルモンおよび性腺刺激ホルモン放出ホルモン投与による卵巣刺激が河川型水牛 (Bubalus bubalis) における経腟採卵-体外受精成績に及ぼす影響2018

    • Author(s)
      坂口謙一郎, Excel Rio S. Maylem, Ramesh C. Tilwani, 栁川洋二郎, 片桐成二, Edwin C. Atabay, Eufrocina P. Atabay, 永野昌志
    • Organizer
      第37回北海道牛受精卵移植研究会研究発表大会
    • Related Report
      2018 Annual Research Report
  • [Presentation] モンゴル国における羊体外受精卵由来産子作出の試み:体外成熟培地への抗酸化剤アスタキサンチンの添加効果2018

    • Author(s)
      永野昌志, Purevdorj Erdenetogtokh, 栁川洋二郎, 片桐成二
    • Organizer
      第37回北海道牛受精卵移植研究会研究発表大会
    • Related Report
      2018 Annual Research Report
  • [Presentation] 液体クロマトグラフィー高分解能質量分析を用いた牛卵子中遊離脂肪酸およびトリアシルグリセロールの分析2018

    • Author(s)
      植芝滉己, 呉ユエ, 陳震, 坂口謙一郎, 柳川洋二郎, 片桐成二, 千葉仁志, 千葉仁志, 惠淑萍, 永野昌志
    • Organizer
      第161回日本獣医学会学術集会
    • Related Report
      2018 Annual Research Report
  • [Presentation] 乳牛の卵巣内胞状卵胞数が血中卵胞刺激ホルモン動態と卵胞における性ステロイドホルモン産生に与える影響2018

    • Author(s)
      坂口謙一郎, 柳川洋二郎, 吉岡耕治, 須田智子, 植芝滉己, 河野光平, 宮本祥代, 片桐成二, 永野昌志
    • Organizer
      第161回日本獣医学会学術集会
    • Related Report
      2018 Annual Research Report
  • [Presentation] Usefulness of gas permeable culture device in in vitro production of competent bovine oocytes by shortened culture period2018

    • Author(s)
      Chelenga Madalitso, Mohammed A. Abdel-Ghani, Kenichiro Sakaguchi, Yojiro Yanagawa, Seiji Katagiri, Masashi Nagano
    • Organizer
      第161回日本獣医学会学術集会
    • Related Report
      2018 Annual Research Report
  • [Presentation] 牛卵子の発生能獲得と卵子体外発育培養に関する研究2018

    • Author(s)
      永野昌志
    • Organizer
      第111回日本繁殖生物学会大会
    • Related Report
      2018 Annual Research Report
    • Invited
  • [Presentation] 牛卵子-顆粒層細胞複合体と内卵胞膜細胞との共培養が卵子の発生能と顆粒層細胞のステロイド合成能に与える影響2017

    • Author(s)
      楊 応華、坂口謙一郎、菅野智裕、栁川洋二郎、片桐成二、永野昌志
    • Organizer
      第36回北海道牛受精卵移植研究会研究発表大会
    • Related Report
      2017 Research-status Report
  • [Presentation] 結過程および液状保存における精子運動性サブポピュレーションの変化2017

    • Author(s)
      菅野智裕、髙橋芳幸、栁川洋二郎、片桐成二、永野昌志
    • Organizer
      第36回北海道牛受精卵移植研究会研究発表大会
    • Related Report
      2017 Research-status Report
  • [Presentation] 牛卵巣の大きさが顆粒層細胞の性ステロイドホルモン産生能および卵子核成熟能に与える影響2017

    • Author(s)
      坂口謙一郎、菅野智裕、楊応華、栁川洋二郎、片桐成二、永野昌志
    • Organizer
      平成29年度日本産業動物獣医学会北海道地区大会
    • Related Report
      2017 Research-status Report
  • [Presentation] 内視鏡を用いたヒグマ(Ursus arctos)における子宮内授精の試み2017

    • Author(s)
      栁川洋二郎、鳥居佳子、菅野智裕、田嶋彩野、冨安洵平、松本直也、坂元秀行、亀井利活、間野勉、片桐成二、永野昌志
    • Organizer
      第23回日本野生動物医学会
    • Related Report
      2017 Research-status Report
  • [Presentation] リカゾウ1個体から採取した精液の輸送後および凍結融解後の精子性状2017

    • Author(s)
      鳥居佳子、菅野智裕、曽地千尋、宗内一平、牛尾佳名子、吉住和規、千田圭、橋本渉、永野昌志、片桐成二、柳川洋二郎
    • Organizer
      第23回日本野生動物医学会
    • Related Report
      2017 Research-status Report
  • [Presentation] Mitochondrial activity and reactive oxygen species (ROS) accumulation in bovine oocytes derived from different culture system for in vitro maturation2017

    • Author(s)
      Purevdorj Erdenetogtokh, Yojiro Yanagawa, Seiji Katagiri, Masashi Nagano
    • Organizer
      第160回日本獣医学会学術集会
    • Related Report
      2017 Research-status Report
  • [Presentation] 精子運動能自動解析装置(CASA)を用いた受精能獲得精子の運動性評価2017

    • Author(s)
      菅野智裕、栁川洋二郎、片桐成二、永野昌志
    • Organizer
      第160回日本獣医学会学術集会
    • Related Report
      2017 Research-status Report
  • [Presentation] ヒグマ精液の凍結保存液に添加する糖の種類および冷却速度が凍結融解後の精液性状に及ぼす影響2017

    • Author(s)
      鳥居佳子、菅野智裕、簑原悠太朗、坂元秀行、松本直也、冨安洵平、松井基純、永野昌志、片桐成二、栁川洋二郎
    • Organizer
      第160回日本獣医学会学術集会
    • Related Report
      2017 Research-status Report
  • [Presentation] 牛卵巣内胞状卵胞数と初期胞状卵胞由来卵子-顆粒層細胞複合体の顆粒層細胞増殖能および卵子発生能の関連性2017

    • Author(s)
      坂口謙一郎、谷田孝志、楊応華、菅野智裕、永井克尚、Mohammed. A. Abdel-Ghani、栁川洋二郎、片桐成二、永野昌志
    • Organizer
      第160回日本獣医学会学術集会
    • Related Report
      2017 Research-status Report
  • [Presentation] Astaxanthin improves the growth parameters, developmental competence, and quality of in vitro grown oocytes derived from bovine early antral follicle2017

    • Author(s)
      Mohammed. A. Abdel-Ghani, Chihiro Kanno, Kenichiro Sakaguchi, Yojiro Yanagawa, Seiji Katagiri, Masashi Nagano
    • Organizer
      第160回日本獣医学会学術集会
    • Related Report
      2017 Research-status Report
  • [Presentation] ニホンザルにおける排卵誘起処置を伴う単回人工授精プログラムの検討2017

    • Author(s)
      栁川洋二郎、菅野智裕、南晶子、兼子明久、印藤頼子、佐藤容、木下こづえ、岡本宗裕、片桐成二、永野昌志
    • Organizer
      第160回日本獣医学会学術集会
    • Related Report
      2017 Research-status Report
  • [Presentation] Relationships between antral follicle count, developmental competence of oocytes, and steroidogenesis of granulosa cells in cattle2017

    • Author(s)
      Kenichiro Sakaguchi, Takashi Tanida, Katsuhisa Nagai, Yinghua Yang, Yojiro Yanagawa, Seiji Katagiri, Masashi Nagano
    • Organizer
      Fourth World Congress of Reproductive Biology
    • Related Report
      2017 Research-status Report 2016 Research-status Report
    • Int'l Joint Research
  • [Presentation] Transition of sperm motility subpopulation in bull’s semen during freezing process in a conventional egg-yolk tris extender evaluated by cluster analysis2017

    • Author(s)
      Chihiro Kanno, Yoshiyuki Takahashi, Yojiro Yanagawa, Seiji Katagiri, Masashi Nagano
    • Organizer
      Fourth World Congress of Reproductive Biology
    • Related Report
      2017 Research-status Report
    • Int'l Joint Research
  • [Presentation] ペレット法により凍結したニホンザル精液に対する融解法の違いが精子性状に与える影響2017

    • Author(s)
      栁川洋二郎, 菅野智裕, 兼子明久, 印藤頼子, 佐藤容, 木下こづえ, 今井啓雄,平井啓久,片桐成二, 永野昌志, 岡本宗裕
    • Organizer
      Cryopreservation Conference 2017
    • Related Report
      2017 Research-status Report
  • [Presentation] Effect of Astaxanthin on developmental competence and apoptosis of oocytes derived from early antral follicles2016

    • Author(s)
      Mohammed. A. Abdel-Ghani, Kenichiro Sakaguchi, Chihiro Kanno, Yinghua Yang, Yojiro Yanagawa, Seiji Katagiri, Masashi Nagano
    • Organizer
      Sapporo Summer Seminor for One Health
    • Place of Presentation
      北海道大学(北海道札幌市)
    • Year and Date
      2016-09-20
    • Related Report
      2016 Research-status Report
    • Int'l Joint Research
  • [Presentation] Effect of prematuration culture duration on developmental competences of bovine small sized oocytes2016

    • Author(s)
      Mohammed. A. Abdel-Ghani, Kenichiro Sakaguchi, Chihiro Kanno, Yinghua Yang, Yojiro Yanagawa, Seiji Katagiri, Masashi Nagano
    • Organizer
      第159回日本獣医学会学術集会
    • Place of Presentation
      日本大学(神奈川県藤沢市)
    • Year and Date
      2016-09-06
    • Related Report
      2016 Research-status Report
  • [Presentation] 牛卵巣内胞状卵胞数と初期胞状卵胞由来卵子顆粒層細胞複合体の性ステロイドホルモン産生能の関係2016

    • Author(s)
      坂口謙一郎、谷田孝志、永井克尚、楊応華、栁川洋二郎、片桐成二、永野昌志
    • Organizer
      第159回日本獣医学会学術集会
    • Place of Presentation
      日本大学(神奈川県藤沢市)
    • Year and Date
      2016-09-06
    • Related Report
      2016 Research-status Report
  • [Presentation] Growth, steroidogenesis and subsequent developmental competence of bovine oocyte-granulosa cell complex co-cultured with theca cells2016

    • Author(s)
      楊応華、菅野智裕、坂口謙一郎、栁川洋二郎、片桐成二、永野昌志
    • Organizer
      第159回日本獣医学会学術集会
    • Place of Presentation
      日本大学(神奈川県藤沢市)
    • Year and Date
      2016-09-06
    • Related Report
      2016 Research-status Report

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Published: 2016-04-21   Modified: 2020-03-30  

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