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Studies on bovine nuclear transfer(cloning)and related technologies

Research Project

Project/Area Number 10556058
Research Category

Grant-in-Aid for Scientific Research (B).

Allocation TypeSingle-year Grants
Section展開研究
Research Field Applied animal science
Research InstitutionHOKKAIDO UNIVERSITY

Principal Investigator

TAKAHASHI Yoshiyuki  Hokkaido University, Graduate School of Veterinary Medicine, Professor, 大学院・獣医学研究科, 教授 (70167485)

Co-Investigator(Kenkyū-buntansha) MORIYASU Satoru  Hokkaido Animal Research Center, Embryo Transfer Laboratory, Researcher, 畜産工学部, 研究員
NAGANO Masashi  Hokkaido University, Graduate School of Veterinary Medicine, Instructor, 大学院・獣医学研究科, 助手 (70312402)
KATAGIRI Seiji  Hokkaido University, Graduate School of Veterinary Medicine, Associate Professor, 大学院・獣医学研究科, 助教授 (00292061)
KISHI Masao  Snow Bland Milk Products Co.Ltd., Embryo Transplantation Laboratory, Researcher, 受精卵移植研究所, 研究員
Project Period (FY) 1998 – 2000
Project Status Completed (Fiscal Year 2000)
Budget Amount *help
¥13,100,000 (Direct Cost: ¥13,100,000)
Fiscal Year 2000: ¥1,900,000 (Direct Cost: ¥1,900,000)
Fiscal Year 1999: ¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1998: ¥9,100,000 (Direct Cost: ¥9,100,000)
KeywordsCattle / Nuclear transfer / Cloning
Research Abstract

Present studies were conducted to establish reliable bovine nuclear transfer(NT)procedure using embryonic blastomeres and somatic cells as nuclear donors, and to develop related technologies such as cryopreservation of oocytes/follicles and in vitro culture of preantral follicles.
In experiments for NT using embryonic blastomeres, we have developed a new oocyte-enucleation technique with a high efficiency by using pre-activated oocytes. Clone calves were obtained after the transfer of NT embryos reconstituted with blastomeres, and we confirmed their consistent growth and fertility. In experiments for the production of clone calves using somatic cells as nuclear doner, we have investigated the several factors affecting the development of NT embryos. Results indicate that cell-cycle coordination between recipient cytoplast and nuclear donors is most critical, and that the combination of nuclear donor in the G0/G1 phase and recipient cytoplast in the M phase produces embryos with high deve … More lopmental capacity when somatic cells are used as nuclear donors. Clone calves were obtained after the transfer of NT embryos reconstructed with uterine epithelial cells, colostrum-derived mammary gland epithelial cells and ear-derived fibroblast cells ; however, abortion after the transfer of NT embryos and abnormalities in the clone calves were frequently observed.
To prepare a large number of recipient oocytes, we have studied to develop a pre-antral follicle culture system and cryopreservation protocol for the cryopreservation of follicles/oocytes using cattle and mouse oocytes and preantral follicles. We confirmed that bovine oocytes derived from early preantral follicles could acquire developmental capacity if they were cultured for more than 8 days before maturation culture. A new vitrification solution(VS), a mixture of ethylene glycol and raffinose was developed after the several investigation using mouse oocytes. Using this new VS, we have succeeded to cryopreserve mouse preantral follicles, and confirmed that the oocytes derived from vitrified-cultured preantral follicles could develop to the blastocyst stage. Less

Report

(4 results)
  • 2000 Annual Research Report   Final Research Report Summary
  • 1999 Annual Research Report
  • 1998 Annual Research Report
  • Research Products

    (37 results)

All Other

All Publications (37 results)

  • [Publications] Mohamed Nour MS: "Preparation of young preactivated oocytes with high enucleation efficiency for bovine nuclear transfer."Theriogenology. 51. 661-666 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] 笹本良彦: "ウシの超音波誘導経膣採卵において卵胞吸引時の吸引針の回転と吸引針の違いが卵子回収率に及ぼす影響."北海道牛受精卵移植研究会報. 18. 19-22 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] 澤井健: "子宮内膜上皮細胞をドナーとした体細胞クローン牛の作出."北海道牛受精卵移植研究会報. 18. 34-37 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Ikeda K: "Effect of medium change on the development of in vitro matured and fertilized bovine oocytes cultured in medium containing amino acids."J Vet Med Sci. 62. 121-123 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Mohamed Nour MS: "In vitro developmental potential of bovine nuclear transfer embryos derived from primary cultured cumulus cells."J Vet Med Sci. 62. 339-342 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Mohamed Nour MS: "Bovine nuclear transfer using cumulus cells derived from serum-starved and confluent cultures."J Reprod Dev. 46. 85-92 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] 永野昌志: "ウシの初期胞状卵胞および一次卵胞の体外培養."北海道牛受精卵移植研究会会報. 19. 8-13 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] 岸昌生: "ポリエチレングリコールおよびジメチルスフオキシドがウシ初乳由来乳腺上皮細胞を用いた核移植胚の融合および体外発育に及ぼす影響."北海道牛受精卵移植研究会会報. 19. 19-23 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Dela Pena EC: "エチレングリコールとラフイノースを用いたマウス卵子のガラス化における前処理の効果."北海道牛受精卵移植研究会会報. 19. 31-34 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Cheong HT: "Development of reconstituted pig embryos by nuclear transfer of cultured cumulus cells."Reprod Fertil Dev. 12. 15-20 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Kishi M: "Nuclear transfer in cattle using colostrumu-derived mammary gland epithellial cells and ear-derived fibroblast cells."Theriogenology. 54. 675-684 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Bishonga C: "In vitro growth of mouse ovarian preantral follicles and the capacity of their oocytes to develop to the blastocyst stage."J Vet Med Sci. 63(in press). (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Mohamed Nour MS, Takahashi Y: "Preparation of young preactivated oocytes with high enucleation efficiency for bovine nuclear transfer."Theriogenology. 51. 661-666 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Sasamoto Y, Katagiri S, Takahashi Y: "Effects of twisting and type of aspiration needles on the efficiency of transvaginal ultrasound-guided transvaginal ovum pick-up in cattle."Bull Soc Hokkaido Bovine ET. 18. 19-22 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Sawai T, Moriyasu S, Kageyama S, Yamamoto Y, Ashino M, Kitano N, Matsuzaki S, Yamashina H, Okamoto H, Hujikawa A, Minamihashi A: "Birth of a clone calf after the transfer of nuclear transfer embryos reconstructed with uterine epithelial cells."Bull Soc Hokkaido Bovine ET. 18. 34-37 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Ikeda K, Takahashi Y, Katagiri S: "Effect of medium change on the development of in vitro matured and fertilized bovine oocytes cultured in medium containing amino acids."J Vet Med Sci. 62. 121-123 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Mohamed Nour MS, Takahashi Y: "In vitro developmental potential of bovine nuclear transfer embryos derived from primary cultured cumulus cells."J Vet Med Sci. 62. 339-342 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Mohamed Nour MS, Ikeda K, Takahashi Y: "Bovine nuclear transfer using cumulus cells derived from serum-starved and confluent cultures."J Reprod Dev. 46. 85-92 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Nagano M, Ikawa M, Yamamoto N, Katagiri S, Takahashi Y: "In vitro culture of bovine preantral and early antral follicles."Bull Soc Hokkiado Bovine ET. 19. 8-13 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Kishi M, Itagaki Y, Takakura R, Sudo T, Yoshinarai M, Kashima N: "Effects of polythylenegycol and DMSO on the fusion of oocytes and their subsequent in vitro development of nuclear transplanted bovine embryos reconstructed with colostrum-derived mammary gland epithelial cells."Bull Soc Hokkaido Bovine ET. 19. 19-23 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Dela Pena EC, Atabay EC, Nagano M, Katagiri S, Takahashi Y: "Effects of pretreatment on the viability of mouse oocytesvitrified with a mixture of ethylene glycol and raffinose."Bull Soc Hokkaido Bovine ET. 19. 31-34 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Cheong HT, Ikeda K, Martinez Diaz MA, Katagiri S, Takahashi Y: "Development of reconstituted pig embryos by nuclear transfer of cultured cumulus cells."Reprod Fertil Dev. 12. 15-20 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Kishi M, Itagaki Y, Takakura R, Imamura M, Sudo T, Yoshinarai M, Tanimoto M, Yasue H, Kashima N: "Nuclear transfer in cattle using colostrumu-derived mammary gland epithelial cells and ear-derived fibroblast cells."Theriogenology. 54. 875-684 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] Bishonga C, Takahashi Y, Katagiri S, Nagano M, Ishikawa A: "In vitro growth of mouse ovarian preantral follicles and the capacity of their oocytes to develop to the blastocyst stage."J Vet Med Sci. 63, (in press). (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2000 Final Research Report Summary
  • [Publications] 永野昌志: "ウシの初期細胞状卵胞および一次卵胞の体外培養."北海道牛受精卵移植研究会会報. 19. 8-13 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] 岸昌生: "ポリエチレングリコールおよびジメチルスフオキシドがウシ初乳由来乳腺上皮細胞を用いた核移植胚の融合および体外発育に及ぼす影響."北海道牛受精卵移植研究会会報. 19. 19-23 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Dela Pena EC: "エチレングリコールとラフイノースを用いたマウス卵子のガラス化における前処理の効果."北海道牛受精卵移植研究会会報. 19. 31-34 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Cheong HT: "Development of reconstituted pig embryos by nuclear transfer of culture cumulus cells."Reprod Fertil Dev. 12. 15-20 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Kishi M: "Nuclear transfer in cattle using colostrumu-derived mammary gland epithellial cells and ear-derived fibroblast cells."Theirogenology. 54. 675-684 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Bishonga C: "In vitro growth of mouse ovarian preantral follicles and the capacity of their oocytes to develop to the blastocyst stage."J Vet Med Sci. 63(in press). (2001)

    • Related Report
      2000 Annual Research Report
  • [Publications] Mohamed Nour MS: "Preparation of youn preactivated oocytes with high enucleation efficiency for bovinenuclear transfer"Theriogenology. 51・3. 661-666 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] 松井 基純: "体外成熟培地へ天下したIGF-Iはウシ卵子の成熟を促進しその後の体外受精率を高める"北海道牛受精卵移植研究会報. 18. 13-18 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] 笹本 良彦: "ウシの超音波誘導経膣採卵において卵胞吸引時の吸引針の回転と吸引針の違いが卵子回収成績に及ぼす影響"北海道牛受精卵移植研究会報. 18. 19-22 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Ikeda, K.: "Effect of medium change on the development of in vitro matured and fertilized bovine oocytes cultured in medium containing amino acids"J Vet Med Sci. 62・1. 121-123 (2000)

    • Related Report
      1999 Annual Research Report
  • [Publications] Mohamed Nour MS: "In vitro developmental potential of bovine nuclear transfer embryos derived from primary cultured cumulus cells"J Vet Med Sci. 62・3. 339-342 (2000)

    • Related Report
      1999 Annual Research Report
  • [Publications] Mohamed Nour MS: "Bovine nuclear transfer using cumulus cells derived from serum-starved and confluent cultures"J Reprod Dev. 46・2(in press). (2000)

    • Related Report
      1999 Annual Research Report
  • [Publications] Mohamed Nour MS: "Preparation of young preactivated oocytes with high enucleation efficiency for bovine nuclear transfer." Theriogenology. 51・3(in press). (1999)

    • Related Report
      1998 Annual Research Report

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Published: 1998-04-01   Modified: 2016-04-21  

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