Establishment of embryonic stem cell lines from in vitro matured and in vitro fertilized dog oocytes and their application to regenerative veterinary medicine

2011 Fiscal Year Final Research Report

• Project/Area Number: 20380174
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Clinical veterinary science
• Research Institution: Osaka Prefecture University
• Principal Investigator: INABA Toshio 大阪府立大学, 生命環境科学研究科, 教授 (00137241)
• Co-Investigator(Kenkyū-buntansha): SHUGIURA Kikuya 大阪府立大学, 生命環境科学研究科, 准教授 (30171143) ; HATOYA Shingo 大阪府立大学, 生命環境科学研究科, 助教 (40453138) ; SAKURAGAWA Norio 北里大学, 医療衛生学部, 研究員 (70183374) ; TORII Ryuzo 滋賀医科大学, 動物生命科学研究センター, 教授 (50106647) ; TAMADA Hiromichi 大阪府立大学, 生命環境科学研究科, 教授 (10155252) ; KAWATE Noritoshi 大阪府立大学, 生命環境科学研究科, 准教授 (80221901) ; KIDA Kayoko 大阪府立大学, 生命環境科学研究科, 助教 (50405362) ; TANI Hiroyuki 大阪府立大, 生命環境科学研究科, 准教授 (00305658)
• Project Period (FY): 2008 – 2011
• Keywords: 獣医学 / トランスレーショナルリサーチ / バイオテクノロジー / 移植・再生医療 / 発生・分化 / ES細胞

Research Abstract

The objective of this study was to establish embryonic stem(ES) cell lines from in vitro matured and fertilized dog oocytes for regenerative veterinary medicine, and the following results were obtained.(1) We successfully enhanced oocyte growth and maturation by controlling the induction of fertile oestrus in immature and dioestrous bitches.(2) The in vitro maturation rate of canine oocytes was improved by embedding and culturing in collagen gels.(3) Canine ES-like cell colonies were formed by culturing inner cell masses collected from blastocysts in uterine horns, and maintained their proliferation and morphology beyond passage 10.(2) Neurons could be induced simply from primate ES cells.(3) Bone marrow stromal cell transplantation caused no complications and could have beneficial effects on functional recovery of dogs with spinal cord injury, and further studies are required to apply canine ES cells for the treatment of these dogs.

Research Products

• [Journal Article] Methods for cell harvesting and the biological properties of canine bone marrow stromal cells at successive passages (2012)
  • Author(s): Nishida H, Shoji Y, Nakamura M, Hatoya S, Sugiura K, Yamate J, Kuwamura M, Kotani T, Nakayama M, Suzuki Y, Ide C, Inaba T
  • Journal Title: Am. J. Vet. Res Volume: (印刷中)
  • DOI: DOI:10.2460/ajvr.72.8.1118
  • Peer Reviewed
• [Journal Article] Safety of autologous bone marrow stromal cell transplantation in dogs with acute spinal cord injury (2012)
  • Author(s): Nishida H, Nakayama M, Tanaka H, Kitamura M, Hatoya S, Sugiura K, Harada Y, Suzuki Y, Ide C, Inaba T
  • Journal Title: Vet. Surg Volume: (印刷中)
  • DOI: DOI:10.1111/j.1532-950X.2011.00959.x
• [Journal Article] Plasma insulin-like peptide3and testosterone concentrations in male dogs : changes with age and effects of cryptorchidism (2012)
  • Author(s): Pathirana IN, Yamasaki H, Kawate N, Tsuji M, Bullesbach EE, Takahashi M, Hatoya S, Inaba T, Tamada H
  • Journal Title: Theriogenology Volume: 77巻 Pages: 550-557
  • DOI: DOI:10.1016/j.theriogenology.2011.08.030
  • Peer Reviewed
• [Journal Article] In vitro effects of estradiol-17β, monobutyl phthalate and mono-(2-ethyl hexyl) phthalate on the secretion of testosterone and insulin-like peptide 3 by interstitial cells of scrotal and retained testes in dogs (2011)
  • Author(s): Pathirana IN, Kawate N, Tsuji M, Takahashi M, Hatoya S, Inaba T, Tamada H
  • Journal Title: Theriogenology Volume: 76巻 Pages: 1227-123 3
  • DOI: DOI:10.1016/j.theriogenology.2011.05.027
  • Peer Reviewed
• [Journal Article] Induction of fertile oestrus in dioestrous bitches by prostaglandin F2α and a GnRH-agonist (2011)
  • Author(s): Ohmura M, Torii R, Hatoya S, Sugiura K, Tamada H, Kawate N, Takahashi M, Sawada T, Inaba T
  • Journal Title: Vet. Rec Volume: 168巻 Pages: 669
  • DOI: DOI:10.1136/vr.d1183
• [Journal Article] Evaluation of transplantation of autologous bone marrow stromal cell into the cerebrospinal fluid for treatment of chronic spinal cord injury in dogs (2011)
  • Author(s): Nishida H, Nakayama M, Tanaka H, Kitamura M, Hatoya S, Sugiura K, Suzuki Y, Ide C, Inaba T
  • Journal Title: Am. J. Vet. Res Volume: 72巻 Pages: 1118-1123
  • DOI: DOI:10.2460/ajvr.72.8.1118
  • Peer Reviewed
• [Journal Article] Comparison of testosterone and insulin-like peptide 3 secretions in response to human chorionic gonadotropin in cultured interstitial cells from scrotal and retained testes in dogs (2011)
  • Author(s): Pathirana IN, Ashida Y, Kawate N, Tanaka K, Makoto T, Takahashi M, Hatoya S, Inaba T, Tamada H
  • Journal Title: Anim. Reprod. Sci Volume: 124巻 Pages: 138-144
  • DOI: DOI:10.1016/j.anireprosci.2011.02.014
  • Peer Reviewed
• [Journal Article] Effect of IL-12 on the canine dendritic cell maturation following differentiation induced by granulocyte-macrophage CSF and IL-4 (2010)
  • Author(s): Sugiura K, Wijewardana V, Fujimoto M, Akazawa T, Yahata M, Mito K, Hatoya S, Inoue N, Inaba T
  • Journal Title: Vet. Immunol. Immunopathol Volume: 137巻 Pages: 322-326
  • DOI: DOI:10.1016/j.vetimm.2010.06.006
  • Peer Reviewed
• [Journal Article] Detection of relaxin mRNA in the corpus luteum, uterus, and uterine cervix in the bitch (2010)
  • Author(s): Tamada H, Kawate N, Kawata N, Inaba T, Kida K, Hatoya S, Akune A, Nakama K, Kohsaka T, Sawada T
  • Journal Title: J. Vet. Med. Sci Volume: 72巻 Pages: 1383-1386
  • DOI: DOI:10.1292/jvms.09-0485
  • Peer Reviewed
• [Journal Article] INFγmarkedly cooperates with intratumoral dendritic cell vaccine in dog tumor models (2010)
  • Author(s): Mito K, Sugiura K, Ueda K, Hori T, Akazawa T, Yamate J, Hatoya S, Inaba M, Inoue N, Ikehara S, Inaba T
  • Journal Title: Cancer Res Volume: 70巻 Pages: 7093-7101
  • DOI: DOI:10.1158/0008-5472.CAN-10-0600
  • Peer Reviewed
• [Journal Article] Analysis of single nucleotide polymorphisms in the 3' region of the estrogen receptor 1 gene in normal and cryptorchid miniature dachshunds and chihuahuas (2010)
  • Author(s): Pathirana IN, Tanaka K, Kawate N, Tsuji M, Kida K, Hatoya S, Inaba T, Tamada H
  • Journal Title: J. Reprod. Dev Volume: 56巻 Pages: 405-410
  • URL: http://dx.doi.org/10.1262/jrd.09-195T
  • Peer Reviewed
• [Journal Article] Self-contained induction of neurons from human embryonic stem cells (2009)
  • Author(s): Okuno T, Nakayama T, Konishi N, Michibata H, Wakimoto K, Suzuki Y, Nito S, Inaba T, Nakano I, Muramatsu S, Takano M, Kondo Y, Inoue N
  • Journal Title: PLoS One Volume: 4巻 Pages: e6318
  • DOI: 10.1371/journal.pone.0006318
  • Peer Reviewed
• [Journal Article] Canine oocyte maturation in culture : significance of estrogen and EGF receptor gene expression in cumulus cells (2009)
  • Author(s): Hatoya S, Sugiyama Y, Nishida H, Okuno T, Torii R, Sugiura K, Kida K, Kawate N, Tamada H, Inaba T
  • Journal Title: Theriogenology Volume: 71巻 Pages: 560-567
  • DOI: DOI:10.1016/j.theriogenology.2008.08.013
  • Peer Reviewed
• [Journal Article] Construction of an expression vector for improved secretion of canine IL-18 (2008)
  • Author(s): Sugiura K, Akazawa T, Fujimoto M, Wijewardana V, Mito K, Hatoya S, Taketani S, Komori M, Inoue N, Inaba T
  • Journal Title: Vet. Immunol. Immunopathol Volume: 126巻 Pages: 388-391
  • DOI: DOI:10.1016/j.vetimm.2008.07.011
  • Peer Reviewed
• [Presentation] ネコ受精卵の体外培養法の改善 (2012)
  • Author(s): 稲葉俊夫
  • Organizer: 第137回日本生殖医学会関西支部集談会
  • Place of Presentation: 大阪
  • Year and Date: 2012-03-03
• [Presentation] イヌES細胞の研究から再生獣医療に向けて (2012)
  • Author(s): 稲葉俊夫
  • Organizer: 第17回滋賀県獣医学会
  • Place of Presentation: 滋賀
  • Year and Date: 2012-02-26
• [Presentation] イヌ骨髄間質細胞の神経幹細胞に及ぼす影響 (2011)
  • Author(s): 稲葉俊夫
  • Organizer: 第152回日本獣医学会学術集会
  • Place of Presentation: 大阪
  • Year and Date: 2011-09-20
• [Presentation] ES細胞の樹立に向けたネコ受精卵の体外培養法に関する研究 (2011)
  • Author(s): 稲葉俊夫
  • Organizer: 第152回日本獣医学会学術集会
  • Place of Presentation: 大阪
  • Year and Date: 2011-09-20
• [Presentation] イヌ顆粒球減少症に向けたイヌ造血因子の作製 (2011)
  • Author(s): 稲葉俊夫
  • Organizer: 第152回日本獣医学会学術集会
  • Place of Presentation: 大阪
  • Year and Date: 2011-09-19
• [Presentation] FSHおよびhCGによるイヌ卵子の体外成熟率の改善 (2011)
  • Author(s): 稲葉俊夫
  • Organizer: 第136回日本生殖医学会関西支部集談会
  • Place of Presentation: 大阪
  • Year and Date: 2011-03-05
• [Presentation] サルES細胞からのオリゴデンドロサイトへの簡便な分化誘導法 (2010)
  • Author(s): 稲葉俊夫
  • Organizer: 第150回日本獣医学会学術集会
  • Place of Presentation: 北海道
  • Year and Date: 2010-09-16
• [Presentation] 再生医療の最前線-幹細胞:犬ES細胞(胚性幹細胞)の開発 (2010)
  • Author(s): 鳩谷晋吾
  • Organizer: 第149回日本獣医学会学術集会
  • Place of Presentation: 東京
  • Year and Date: 2010-03-26
• [Presentation] ES細胞によるパーキンソン病モデル動物の治療に関する基礎的研究 (2010)
  • Author(s): 稲葉俊夫
  • Organizer: 第149回日本獣医学会学術集会
  • Place of Presentation: 東京
  • Year and Date: 2010-03-26
• [Presentation] イヌ骨髄間質細胞の継代に伴う特性変化 (2009)
  • Author(s): 稲葉俊夫
  • Organizer: 第148回日本獣医学会学術集会
  • Place of Presentation: 鳥取
  • Year and Date: 2009-09-27
• [Presentation] コラーゲン・ゲルを用いたイヌ卵母細胞の体外発育 (2009)
  • Author(s): 稲葉俊夫
  • Organizer: 第134回日本生殖医学会関西支部集談会
  • Place of Presentation: 大阪
  • Year and Date: 2009-03-14
• [Presentation] コラーゲン・ゲル培養イヌ卵母細胞の体外発育に及ぼすhypoxanthineとFSHの影響 (2008)
  • Author(s): 稲葉俊夫
  • Organizer: 第146回日本獣医学会学術集会
  • Place of Presentation: 宮崎
  • Year and Date: 2008-09-24
• [Remarks]
  • URL: http://kyoindb.acs.osakafu-u.ac.jp/profile/out.cudjDYHfHMO.PEVGDxV-iw==.html
• [Patent(Industrial Property Rights)] 腫瘍治療用組成物およびその応用 (1998)
  • Inventor(s): 杉浦喜久弥、水戸凱、稲葉俊夫
  • Industrial Property Rights Holder: 公立大学法人大阪府立大学
  • Industrial Property Number: 特許特願2008-143 465号、特開2009-13166号
  • Filing Date: 1998-05-30