Development of chicken primordial germ cell manipulation methods and its application to production of transgenic chickens

2010 Fiscal Year Self-evaluation Report

• Project/Area Number: 20380156
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Applied animal science
• Research Institution: National Institute of Agrobiological Sciences
• Principal Investigator: NAITO Mitsuru National Institute of Agrobiological Sciences, 遺伝子組換え家畜研究センター, 上級研究員 (70355733)
• Project Period (FY): 2008 – 2012
• Keywords: 始原生殖細胞 / 外来遺伝子 / トランスフェクション / GFP遺伝子 / ニワトリ初期胚 / 生殖系列キメラ / 品種識別 / 体外培養

Research Abstract

形質転換ニワトリ作出技術の開発は、卵白中への医薬品等の有用物質の生産、遺伝子機能の解析、さらにはニワトリ集団の遺伝的改良など、多方面への応用が期待されている。本研究では、始原生殖細胞の培養条件を開発・改良するため、フィーダー細胞の調整法の検討、フィーダー細胞上での始原生殖細胞の維持・増殖のための培養液の組成の改良、始原生殖細胞への遺伝子導入法の検討、遺伝子導入された始原生殖細胞の薬剤による選択条件の解明、等を行う。そして、外来遺伝子が導入された始原生殖細胞をレシピエント胚に移植して、生殖系列キメラニワトリを介して個体に再生するシステムを構築する。以上のシステムを用いて、ニワトリ個体への外来遺伝子導入技術を開発する。

Research Products

• [Journal Article] In vitro culture of testicular and ovarian gonocytes obtained from 19-day incubated chicken embryos and subsequent colonization into gonads of recipient embryos. (2011)
  • Author(s): Naito M, Harumi T, Kuwana T
  • Journal Title: Journal of Poultry Science 48 Pages: 112-118
  • Peer Reviewed
• [Journal Article] Long term in vitro culture of chicken primordial germ cells isolated from embryonic blood and incorporation into germline of recipient embryo. (2010)
  • Author(s): Naito M, Harumi T, Kuwana T
  • Journal Title: Journal of Poultry Science 47 Pages: 57-64
  • Peer Reviewed
• [Journal Article] Preferential migration of transferred primordial germ cells to left germinal ridge of recipient embryos in chickens. (2009)
  • Author(s): Naito M, Minematsu T, Harumi T, Kuwana T
  • Journal Title: Journal of Poultry Science 46 Pages: 40-45
  • Peer Reviewed
• [Journal Article] Quantitative genotyping by amplifying the polymorphic sequences of Pre-Melanosomal Protein (PMEL17) gene using real-time polymerase chain reaction in chickens. (2008)
  • Author(s): Minematsu T, Harumi T, Naito M
  • Journal Title: British Poultry Science 49 Pages: 542-549
  • Peer Reviewed
• [Journal Article] Germ cell-specific expression of GFP gene induced by chicken vasa homologue (Cvh) promoter in early chicken embryos. (2008)
  • Author(s): Minematsu T, Harumi T, Naito M
  • Journal Title: Molecular Reproduction and Development 75 Pages: 1515-1522
  • Peer Reviewed
• [Presentation] Spermatogonial stem cells. (2011)
  • Author(s): 内藤充
  • Organizer: First International Symposium on Propagation of Endangered Species of Birds.
  • Place of Presentation: Abu Dhabi, United Arab Emirates.
  • Year and Date: 20110208-20110210
• [Presentation] In vitro culture of chicken primordial germ cells isolated from embryonic blood. (2010)
  • Author(s): Naito M, Harumi T, Kuwana T.
  • Organizer: XIIIth European Poultry Conference
  • Place of Presentation: Tours, France.
  • Year and Date: 20100823-20100827
• [Presentation] Basic studies for transgenic chickens. (2010)
  • Author(s): 内藤充.
  • Organizer: International Workshop on Preservation of Avian Primordial Germ Cells and Its Usage.
  • Place of Presentation: Okinawa, Japan.
  • Year and Date: 20100126-20100128
• [Presentation] ニワトリ初期胚血液より採取した始原生殖細胞培養の試み (2010)
  • Author(s): 内藤充・春海隆・桑名貴
  • Organizer: 日本家禽学会2010年度春季大会
  • Place of Presentation: 明治大学駿河台キャンパス、東京都千代田区
  • Year and Date: 2010-03-30
• [Presentation] 始原生殖細胞を利用した鳥類の生殖制御法 (2009)
  • Author(s): 内藤充
  • Organizer: 第102回日本繁殖生物学会大会
  • Place of Presentation: 近畿大学農学部、奈良県奈良市
  • Year and Date: 20090926-20090927
• [Book] 畜産技術発達史 12.鶏の人工孵化と雌雄鑑別技術(分担執筆)(畜産技術協会)(東京) (2011)
  • Author(s): 内藤充
  • Total Pages: 106-113
• [Book] 畜産技術発達史 35.採卵鶏の光線管理技術(分担執筆)(畜産技術協会)(東京) (2011)
  • Author(s): 内藤充
  • Total Pages: 240-245
• [Book] 日本農学80年史第33章家禽学 3.発生工学 (分担執筆) (2009)
  • Author(s): 内藤 充
  • Total Pages: 238
  • Publisher: 養賢堂