Genetic analysis of avian stem cells and the application for poultry breeding

• Project/Area Number: 26450376
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Animal production science
• Research Institution: Shinshu University
• Principal Investigator: Kagami Hiroshi 信州大学, 学術研究院農学系, 教授 (80308303)
• Co-Investigator(Kenkyū-buntansha): 小野 珠乙 信州大学, 学術研究院農学系, 教授 (10177264) ; 鈴木 俊介 信州大学, 学術研究院農学系, 助教 (30431951)
• Project Period (FY): 2014-04-01 – 2018-03-31
• Project Status: Completed (Fiscal Year 2017)
• Budget Amount: ¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000); Fiscal Year 2017: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2016: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2015: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2014: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
• Keywords: ニワトリ / 幹細胞 / 始原生殖細胞 / 遺伝的特徴 / 生殖キメラ / 細胞育種 / 家禽 / 遺伝的特性解析 / 発生分化 / 再生 / ゲノム編集

Outline of Final Research Achievements

Regeneration of tissues and organs by stem cell differentiation has hardly been achieved in poultry. In the present studies, stem cells from the early embryo or the primordial germ cells (PGCs) from the circulating blood of the early embryos were isolated and the developmental mechanism was analyzed. These stem cells and primordial germ cells were used as donor and transplanted into the recipient. Donor stem cells derived from chicken early embryo can differentiate into various cell lineages including germ line. It was also revealed that germline chimeras can be efficiently produced using the primordial germ cells in early embryonic circulation. Furthermore, we cultivated efficient culturing methods of primordial germ cells. Genomic editing was performed on these cultured primordial germ cells and knockout of the ovomucoid control gene in egg white was performed for the first time.
These results seemed to contribute to innovation of cell breeding in poultry in the future.

Research Products

• [Journal Article] Autonomous xenogenic cell fusion of murine and chick skeletal muscles myoblasts (2017)
  • Author(s): Takaya, T., Nihashi, Y., Kojima, S., Ono, T. and Kagami, H.
  • Journal Title: Animal Science Journal Volume: 88 Issue: 11 Pages: 1880-1885
  • DOI: 10.1111/asj.12884
  • NAID: 120007099848
  • Peer Reviewed / Open Access
• [Journal Article] Perspectives on Avian Stem Cells for Poultry Breeding (2016)
  • Author(s): Hiroshi Kagami
  • Journal Title: Animal Science Journal Volume: 87 Issue: 9 Pages: 1065-1075
  • DOI: 10.1111/asj.12620
  • NAID: 120007099644
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Chicken Stem Cell Factor Enhances Primordial Germ Cell Proliferation Cooperatively with Fibroblast Growth Factor 2 (2016)
  • Author(s): Daichi Miyahara, Isao Oishi, Ryuichi Makino, Nozomi Kurumisawa, Ryuma Nakaya, Tamao Ono, Hiroshi Kagami, Takahiro Tagami.
  • Journal Title: The Journal of Reproduction and Development Volume: 62 Pages: 143-149
  • NAID: 130005146458
  • Peer Reviewed / Open Access
• [Journal Article] Targeted mutagenesis in chicken using CRISPR/Cas9 system (2016)
  • Author(s): Oishi I, Yoshii K, Miyahara D, Kagami H, Tagami T
  • Journal Title: Scientific Reports Volume: 6 Issue: 1 Pages: 23980-23980
  • DOI: 10.1038/srep23980
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Stem Cell Application for Poultry Breeding. (2015)
  • Author(s): Hiroshi Kagami
  • Journal Title: Proceedings of the International Symposium on Frontier Biotechnology 2015 & 2nd International Symposium, Institute for Biomedical Sciences. Volume: 1 Pages: 7-8
  • Peer Reviewed
• [Journal Article] Culture Conditions for Maintain Propagation, Long-term Survival and Germline Transmission of Chicken Primordial Germ Cell-Like Cells (2014)
  • Author(s): Daichi Miyahara, Takafumi Mori, Ryuichi Makino, Yoshiaki Nakamura, Isao Oishi, Tamao Ono, Keijiro Nirasawa, Takahiro Tagami, Hiroshi Kagami
  • Journal Title: The Journal of Poultry Science Volume: 51 Issue: 1 Pages: 87-95
  • DOI: 10.2141/jpsa.0130077
  • NAID: 130004446431
  • ISSN: 1346-7395, 1349-0486
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Presentation] Avian Stem Cells for Regenerative Studies & Germplasm Conservation (2017)
  • Author(s): Hiroshi Kagami
  • Organizer: International Forum on Avian Germplasm and Genome Editing
  • Invited
• [Presentation] Stem Cell Application for Poultry Breeding (2016)
  • Author(s): Hiroshi Kagami
  • Organizer: The 17th Asian-Australasian Association of Animal Production Societies Animal Science Congress
  • Place of Presentation: Kyushu Sangyo University, Fukuoka
  • Year and Date: 2016-08-24
  • Int'l Joint Research / Invited
• [Presentation] ゲノム編集技術による卵白アレルゲン「オボムコイド」ノックアウトニワトリの作出 (2016)
  • Author(s): 大石勳、吉井京子、宮原大地、鏡味裕、田上貴寛
  • Organizer: 日本家禽学会2016年度春季大会
  • Place of Presentation: 日本獣医生命科学大学（東京都内）
  • Year and Date: 2016-03-30
• [Presentation] 発生工学の新展開 (2015)
  • Author(s): 鏡味裕
  • Organizer: 第38回日本分子生物学会年会・第88回日本生化学会大会合同大会
  • Place of Presentation: 神戸国際会議場（神戸市内）
  • Year and Date: 2015-12-01
  • Invited
• [Presentation] 幹細胞キメラ作出におけるユバ粕給与が及ぼす影響 (2015)
  • Author(s): 鏡味裕、安藤貞、今泉勇作、石川美佳、田村芽衣、大石勳、橋谷田豊
  • Organizer: 日本畜産学会第119回大会
  • Place of Presentation: 宇都宮大学（宇都宮市）
  • Year and Date: 2015-03-28 – 2015-03-30
• [Presentation] ニワトリ始原生殖細胞における馴化培地・フィーダー細胞フリー培養系の改良 (2014)
  • Author(s): 宮原大地、大石勳、牧野龍一、小野珠乙、鏡味裕、田上貴寛
  • Organizer: 第37回日本分子生物学会年会
  • Place of Presentation: パシフィコ横浜（横浜市）
  • Year and Date: 2014-11-25 – 2014-11-27