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Establishment of embryonic stem cell lines from the dog and their application to regenerative veterinary medicine

Research Project

Project/Area Number 14360191
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Applied veterinary science
Research InstitutionOsaka Prefecture University

Principal Investigator

INABA Toshio  Osaka Prefecture University, Graduate School of Life and Environmental Sciences, Professor, 生命環境科学研究科, 教授 (00137241)

Co-Investigator(Kenkyū-buntansha) SUGIURA Kikuya  Osaka Prefecture University, Graduate Sch. of Life and Environ.Sci., Associate Prof., 生命環境科学研究科, 助教授 (30171143)
KUMAGAI Daijiro  Osaka Prefecture University, Graduate Sch. of Life and Environ.Sci., Research Assoc., 生命環境科学研究科, 助手 (70316016)
TANI Hiroyuki  Osaka Prefecture University, Graduate Sch. of Life and Environ.Sci., Research Assoc., 生命環境科学研究科, 助手 (00305658)
TORII Ryuzo  Siga University of Medical Science, Research Center for Animal Life Science, Professor, 動物生命科学研究センター, 教授 (50106647)
Project Period (FY) 2002 – 2005
Project Status Completed (Fiscal Year 2005)
Budget Amount *help
¥14,800,000 (Direct Cost: ¥14,800,000)
Fiscal Year 2005: ¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 2004: ¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 2003: ¥4,500,000 (Direct Cost: ¥4,500,000)
Fiscal Year 2002: ¥5,900,000 (Direct Cost: ¥5,900,000)
Keywordsdog / embryonic stem cell / oocyte / in vitro maturation / in vitro fertilization / embryonic fibroblast cell / estrogen / estrus induction / 上皮成長因子 / 精子 / EGF / MHC欠損マウス / 胚性幹細胞 / 卵丘細胞 / エストロジェンレセプター遺伝子 / 性腺刺激ホルモン放出ホルモン / アロマターゼ遺伝子
Research Abstract

Embryonic stem (ES) cells are pluripotent cells with the capacity to generate any type of cell. The objective of this study was to establish ES cell lines from the dog for regenerative veterinary medicine, and the following results were obtained.
1.The rise in the levels of hypothalamic P450 aromatase mRNA, hypothalamic and pituitary estrogen receptor α and β mRNAs, and plasma estradiol-17β was associated with termination of anestrus in the bitch.
2.The intranasal administration of a depot-injectable form of the potent GnRH agonist leuprolide acetate was a useful method for controlling the induction of estrus in anestrous bitches that appears to cause the animals little stress, and a combination of leuprolide acetate and oral estradiol-17β might be more effective in inducing estrus.
3.Although the in vitro maturation rate of canine MI oocytes cultured with estrogen and epidermal growth factor was higher than for oocytes cultured in culture media only, the rate that reached the MII stage w … More as not different in both groups.
4.Co-culture of embryonic fibroblast cells enhanced nuclear and cytoplasmic maturation of canine oocytes, and co-culture with canine embryonic fibroblasts developed the embryo up to the 16-cell stage, and with mouse embryonic fibroblasts up to morula stage.
5.Primary canine ES-like cell colonies were formed in 67.9% of inner cell masses from hatched blastocysts, which was significantly higher than from blastocysts (25.6%) and d morulae stage (0%).
6.Canine ES-like cells had typical stem cell-like morphology, expressed specific markers such as alkaline phosphatase activity, stage specific embryonic antigen-1 and Oct-4, and formed embryoid bodies (EBs) in a suspension culture ; extended culture of EBs resulted in the formation of cystic EBs. When the simple Ms were cultured in tissue culture plates, they differentiated into several types of cells including neuron-like, epithelium-like, fibroblast-like, melanocyte-like and myocardium-like cells.
7.Dissociation of canine ES-like cell colonies with collagenase resulted in immediate differentiation, but with mechanical disaggregation these cells remain undifferentiated, and two ES-like cell lines passed 8 passages. Less

Report

(5 results)
  • 2005 Annual Research Report   Final Research Report Summary
  • 2004 Annual Research Report
  • 2003 Annual Research Report
  • 2002 Annual Research Report
  • Research Products

    (37 results)

All 2006 2005 2004 2003 2002 Other

All Journal Article (29 results) Patent(Industrial Property Rights) (3 results) Publications (5 results)

  • [Journal Article] Isolation and characterization of embryonic stem-like cells from canine blastocysts2006

    • Author(s)
      Shingo Hatoya et al.
    • Journal Title

      Molecular Reproduction and Development 73・3

      Pages: 298-305

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Annual Research Report 2005 Final Research Report Summary
  • [Journal Article] Induction of fertile estrus in bitches by an intranasal spray of GnRH-aognist2006

    • Author(s)
      Shingo Hatoya et al.
    • Journal Title

      The Veterinary Record 158・11

      Pages: 378-379

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Annual Research Report 2005 Final Research Report Summary
  • [Journal Article] Reduction of mucin-1 gene expression associated with increased Escherichia coll adherence in the canine uterus in the early stage of dioestrus2006

    • Author(s)
      Kayoko Ishiguro et al.
    • Journal Title

      The Veterinary Journal (印刷中)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Effect of co-culturing with embryonic fibroblasts on IVM, IVF and IVC of canine oocytes2006

    • Author(s)
      Shingo Hatoya et al.
    • Journal Title

      Theriogenology (印刷中)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Isolation and characterization of embryonic stem-like cells from canine blastocysts2006

    • Author(s)
      Shingo Hatoya et al.
    • Journal Title

      Molecular Reproduction and Development 73:3

      Pages: 298-305

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Induction of fertile estrus in bitches by an intranasal spray of GnRH-agonist2006

    • Author(s)
      Shingo Hatoya et al.
    • Journal Title

      The Veterinary Record 158

      Pages: 378-379

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Reduction of mucin-1 gene expression associated with increased Escherichia coli adherence in the canine uterus in the early stage of dioestrus2006

    • Author(s)
      Kayoko Ishiguro et al.
    • Journal Title

      The Veterinary Journal (in press)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Effect of co-culturing with embryonic fibroblasts on IVM, IVF and IVC of canine oocytes2006

    • Author(s)
      Shingo Hatoya et al.
    • Journal Title

      Theriogenology (in press)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Reduction of mucin-1 gene expression associated with increased Escherichia coli adherence in the canine uterus in the early stage of dioestrus2006

    • Author(s)
      Kayoko Ishiguro et al.
    • Journal Title

      The Veterinary Journal 印刷中

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Effect of co-culturing with embryonic fibroblasts on IVM, IVF and IVC of canine oocytes2006

    • Author(s)
      Shingo Hatoya et al.
    • Journal Title

      Theriogenology 印刷中

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Adenomyosis with severe inflammation in the uterine cervix in a dog2005

    • Author(s)
      Hiromichi Tamada et al.
    • Journal Title

      The Canadian Veterinary Journal 46

      Pages: 333-334

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Annual Research Report 2005 Final Research Report Summary
  • [Journal Article] Detection of transforming growth factor- α and epidermal growth factor receptor mRNA and immunohistochemical localization of the corresponding proteins in the canine uterus during the estrous cycle.2005

    • Author(s)
      Hiromichi Tamada et al.
    • Journal Title

      Histology and Histopathology 7・11

      Pages: 817-824

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Detection of transforming growth factor-α and epidermal growth factor receptor mRNA and immunohistochemical localization of the corresponding proteins in the canine uterus during the estrous cycle2005

    • Author(s)
      Hiromichi Tamada et al.
    • Journal Title

      Histology and Histopathology 7:11

      Pages: 817-824

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Detection of transforming growth factor-α and epidermal growth factor receptor mRNA and immunohistochemical localization of the corresponding proteins in the canine uterus during the estrous cycle2005

    • Author(s)
      Hiromichi Tamada et al.
    • Journal Title

      Histology and Histopathology 7・11

      Pages: 817-824

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Induction of fertile estrus in bitches by an intranasal spray of GnRH-aognist2005

    • Author(s)
      Shingo Hatoya et al.
    • Journal Title

      The Veterinary Record (印刷中)

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Adenomyosis with severe inflammation in the uterine cervix in a dog2005

    • Author(s)
      Hiromichi Tamada et al.
    • Journal Title

      The Canadian Veterinary Journal (印刷中)

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Detection of transforming growth factor-α and epidermal growth factor receptor mRNA and immunohistochemical localization of the corresponding proteins in the canine uterus during the estrous cycle2005

    • Author(s)
      Hiromichi Tamada et al.
    • Journal Title

      Histology and Histopathology (印刷中)

    • Related Report
      2004 Annual Research Report
  • [Journal Article] The effects of the aromatase inhibitor fadrozole hydrochloride on fetuses and uteri in late pregnant rats2004

    • Author(s)
      Hiromichi Tamada et al.
    • Journal Title

      Journal of Endocrinology 180・2

      Pages: 337-345

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Effect of ovarian hormones on periodical changes in immune resistance associated with estrous cycle in the beagle bitch2004

    • Author(s)
      Kikuya Sugiura et al.
    • Journal Title

      Immunobiology 209・8

      Pages: 619-627

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary 2004 Annual Research Report
  • [Journal Article] The effects of the aromatase inhibitor fadrozole hydrochloride on fetuses and uteri in late pregnant rats2004

    • Author(s)
      Hiromichi Tamada et al.
    • Journal Title

      Journal of Endocrinology 180:2

      Pages: 337-345

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Effect of ovarian hormones on periodical changes in immune resistance associated with estrous cycle in the beagle bitch2004

    • Author(s)
      Kikuya Sugiura et al.
    • Journal Title

      Immunobiology 209:8

      Pages: 619-627

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Expression of estrogen receptor α and β genes in the mediobasal hypothalamus, pituitary and ovary during the canine estrous cycle2003

    • Author(s)
      Shingo Hatoya et al.
    • Journal Title

      Neuroscience Letters 347・2

      Pages: 131-135

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Long-term prevention of estrus in the bitch and queen using chlormadinone acetate2003

    • Author(s)
      Hiromichi Tamada et al.
    • Journal Title

      The Canadian Veterinary Journal 44・5

      Pages: 416-417

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Delayed implantation induced by fadrozole hyrochioride in the rats2003

    • Author(s)
      Hiromichi Tamada et al.
    • Journal Title

      Contraception 68・1

      Pages: 65-68

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Expression of estrogen receptor α and β genes in the mediobasal hypothalamus, pituitary and ovary during the canine estrous cycle2003

    • Author(s)
      Shingo Hatoya et al.
    • Journal Title

      Neuroscience Letters 347:2

      Pages: 131-135

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Long-term prevention of estrus in the bitch and queen using chlormadinone acetate2003

    • Author(s)
      Hiromichi Tamada et al.
    • Journal Title

      The Canadian Veterinary Journal 44:5

      Pages: 416-417

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Delayed implantation induced by fadrozole hydrochloride in the rats2003

    • Author(s)
      Hiromichi Tamada et al.
    • Journal Title

      Contraception 68:1

      Pages: 65-68

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Enhancement of aromatase gene expression in the mediobasal hypothalamus during anestrus in the beagle bitch2002

    • Author(s)
      Toshio Inaba et al.
    • Journal Title

      Neuroscience Letters 333・2

      Pages: 107-110

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Enhancement of aromatase gene expression in the mediobasal hypothalamus during anestrus in the beagle bitch2002

    • Author(s)
      Toshio Inaba et al.
    • Journal Title

      Neuroscience Letters 333:2

      Pages: 107-110

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Patent(Industrial Property Rights)] イヌの造血前駆細胞を認識するモノクローナル抗体の製造方法2006

    • Inventor(s)
      杉浦 喜久弥, 井上 ひとみ, 尾市 朋子, 稲葉 俊夫
    • Industrial Property Rights Holder
      公立大学法人大阪府立大学
    • Industrial Property Number
      2006-070915
    • Filing Date
      2006-03-15
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Patent(Industrial Property Rights)] イヌ由来胚性幹細胞、その生産方法、及びその継代培養方法2005

    • Inventor(s)
      稲葉 俊夫, 鳩谷 晋吾, 鳥居 隆三, 杉浦 喜久弥, 熊谷 大二郎
    • Industrial Property Rights Holder
      公立大学法人大阪府立大学
    • Industrial Property Number
      2005-225209
    • Filing Date
      2005-08-03
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Annual Research Report 2005 Final Research Report Summary
  • [Patent(Industrial Property Rights)] イヌ科動物の樹状細胞の分化誘導方法2005

    • Inventor(s)
      杉浦 喜久弥, 稲葉 俊夫, 熊谷 大二郎
    • Industrial Property Rights Holder
      公立大学法人大阪府立大学
    • Industrial Property Number
      2005-229118
    • Filing Date
      2005-08-08
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Publications] Shingo Hatoya et al.: "Expression of estrogen receptor α and β genes in the mediobasal hypothalamus, pituitary and ovary during the canine estrous cycle"Neuroscience Letters. 347・2. 131-135 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Hiromichi Tamada et al.: "Long-term prevention of estrus in the bitch and queen using chlormadinone acetate"The Canadian Veterinary Journal. 44・5. 416-417 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Hiromichi Tamada et al.: "Delayed implantation induced by fadrozole hyrochloride in the rats"Contraception. 68・1. 65-68 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Hiromichi Tamada et al.: "The effects of the aromatase inhibitor fadrozole hydrochloride on fetuses and uteri in late pregnant rats"Journal of Endocrinology. 180・2. 337-345 (2004)

    • Related Report
      2003 Annual Research Report
  • [Publications] Toshio Inaba: "Enhancement of aromatase gene expression in the mediobasal hypothalamus during anestrus in the beagle bitch"Neuroscience Letters. 333・2. 107-110 (2002)

    • Related Report
      2002 Annual Research Report

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Published: 2002-04-01   Modified: 2016-04-21  

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