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Derivation of primordial germ cell lines and their application to transgenic technology

Research Project

Project/Area Number 05454649
Research Category

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

Allocation TypeSingle-year Grants
Research Field Developmental biology
Research InstitutionInstitute of Development, Aging and Caner, Tohoku University

Principal Investigator

MATSUI Yasuhisa  Tohoku University, IDAC,Associate Professor, 加齢医学研究所, 助教授 (40241575)

Co-Investigator(Kenkyū-buntansha) YANAI Nobuaki  Tohoku University, IDAC,Instractor, 加齢医学研究所, 助手 (80200525)
Project Period (FY) 1993 – 1994
Project Status Completed (Fiscal Year 1994)
Budget Amount *help
¥6,100,000 (Direct Cost: ¥6,100,000)
Fiscal Year 1994: ¥2,300,000 (Direct Cost: ¥2,300,000)
Fiscal Year 1993: ¥3,800,000 (Direct Cost: ¥3,800,000)
Keywordsprimordial germ cell / embryonic stem cell / gametogenesis / transgenic mouse / c-Kit / SV40 large-T antigen / トランスシェニックマウス / SV40ラージ-T抗原 / 配偶子 / 増殖因子 / e-Kit / steel因子 / SV40ラージT抗原
Research Abstract

Murine primordial germ cells (PGCs) cultured with membrane associated Steel factor, LIF and bFGF develop to apluripotent cell line, EG cell. Although it contributes to all tissues including germ cell in chimaeric mice, it has not been known whether it can directly develop to germ cells. The aim of this research project is to establish a PGC line that directly differetiates to functional gamete. We have established EG cells from various inbred mice and have found that some cell lines from DBA/2 mouse formed scatter type colonies resembling those of primary cultured PGCs. These cell lines seemed to sustain motility that id a character of PGCs, we next tested their ability to develop to gamates by transplantation of reconstituted gonads to adult testis in which exogenous PGCs differentiate to gametes. A EG cell line derived from DBA/2 mice was reaggregate with somatic cells of 13.5 dpc male genital ridges in matrigel, then the aggregates were transplanted to germ cell-deficient W/W^Vtestes. After 2 months, the testes were recovered and were histologically examined using germ cell-specific TRA98 monoclonal antibody. The EG cell-transplanted testis showed some TRA98 positive cells inside seminiferous tubules, but testes transplanted somatic cells alone did not. This result suggests that the EG cell differentiates to germ cells in testis.
As an alternative approach, we planed to make PGC lines from transgenic mice that overexpress c-Kit in PGCs. We got several transgenic lines with germ line-specific oct-3 gene promotor/c-kit chimaeric gene and some of which express the transgene in testis. preliminary results from primary culture experiments suggest that PGCs from the transgenic mice grow more rapidly or longer than those of normal mice. We will establish PGC lines from them and examine their ability to differentiate to gametes.

Report

(3 results)
  • 1994 Annual Research Report   Final Research Report Summary
  • 1993 Annual Research Report
  • Research Products

    (35 results)

All Other

All Publications (35 results)

  • [Publications] Pierce,D.F.: "Inhibition of mammary duct developmeut but not alveolar out grawth during pregnancy in MMTV-TGFβ1^<S223/225> fransgenic mice" Genes & Development. 7. 2308-2317 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Coffey,R.J.: "Acceleration of mammary neoplasia in transforming grouth factor α transgenic mice by 7,1-dimethyl benzanthracene.21GC02:Cancer Research" 54. 1678-1683 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Terajima,M.: "Stractural organizationof the mouse glycophorin A gene." J.Biochew.116. 1105-1110 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Kato,S.: "Molecular Cloning and expression of mause Mg^<2+>-dependeut protein phosphatase B-4(type2CB-4)." Arch.Biochew.Biophys.in press. (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Matsubara,N.: "Murine polo like kinase 1 gene is expressed in meiotic testicular gerw cells and oocytes." Mol.Reprod.Dev.in press. (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Yarai,N.: "Apoptosis is induced at non-permissive tewperature by a transieut increase in p53 in cell lines immortalized with SV40 T-antigen gene." Exp.Cell,Res.211. 296-300 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] 松居靖久: "最新.細胞内シグナル伝達ライブラリー" 羊土社(印刷中), (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] 松居靖久: "生殖細胞-基礎から最前線まで(岡田益吉,長濱嘉孝編)" 共立出版 (印刷中), (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Pierce, D.F., Jhonston, M.D., Matsui, Y., Robinson, S.D., Gold, L.I., Purchio, A,F., Daniel, C.W., Hogan, B.L.M., and Moses, H.L.: "Inhibition of mammary duct development but not alveolar outgrowth during pregnancy in MMTV-TGFbeta1^<s223/225>transgenic mice." Genes & Development. 7. 2308-2317 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Coffey, R.J., Meise, K.S., Matsui, Y., Hogan, B.L.M., Dempsey, P.J., and Halter, S.A.: "Acceleration of mammary neoplasia in transforming growth factor alpha transgenic mice by 7,12-dimethylbenzanthracene." Cancer Research. 54. 1678-1683 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Terajima, M., Matsui, Y., Copeland, N.G., Gilbert, D.J., Jenkins, N.A., and Obinata, M.: "Structural organization of the mouse glycophorin A gene." J.Biochem.116. 1105-1110 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Kato, S., Tetasawa, T., Kobayashi, T., Ohnishi, M., Sasahara, Y., Kusuda, K., Yanagawa, Y., Hiraga, A., Matsui, Y., and Tamura, S.: "Molecular cloning and expression of mouse Mg^<2+>-dependent protein phosphatase beta-4 (type2Cbeta-4)." Arch.Biochem.Biophys.(in press.).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Matsubara, N., Yanagisawa, M., Nishimune, Y., Obinata, M., and Matsui, Y.: "Murine polo like kinase 1 gene is expressed in meiotic testicular germ cell and oocytes." Mol.Reprod.Dev.(in press.).

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Yanai, N., Obinata, M.: "Apoptosis is induced at nonpermissive temperaure by a transient increase in p53 in cell lines immortalized with ts SV40 T-antigen gene" Exp.cell Res.211. 296-300 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Pierce,D.F.: "Inhibition of mammary duct development but not alveolar out growth during pregrancy in MMTV-TαFβl s223/225 transgenic mice." Genes & Development. 7. 2308-2317 (1993)

    • Related Report
      1994 Annual Research Report
  • [Publications] Coffey,R.J.: "Acceleration of mammary neoplasia in transforming growth factorα transgenic mice by 7,1-dimethyl benzanthracene." Cancer Research. 54. 1678-1683 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Terajima,M.: "Stractural orgunization of the mause glycophavin A gene." J.Biochem.116. 1105-1110 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Kato,S.: "Molecular Cloning and expression of mause Mg^<2+>-dependent protein phosphatase β-4(type 2Cβ-4)." Arch.Biochem.Biophys.in press. (1995)

    • Related Report
      1994 Annual Research Report
  • [Publications] Matsubara,N.: "Murine pdo like kinase l gene is expressed in meiotic testicular genu cells and oocytes." Mol.Reprod.Dev.in press. (1995)

    • Related Report
      1994 Annual Research Report
  • [Publications] Yanai,N.: "Apoptosis is induced at non-permissive tewperature by a transieut increase in p53 in cell lines immortalized with TS-SV40 T-antigen gene." Exp.Cell.Res.211. 296-300 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] 松居靖久: "最新・細胞内シグナル伝達ライブラリー(山本雅編)" 羊土社(印刷中), (1995)

    • Related Report
      1994 Annual Research Report
  • [Publications] 松居靖久: "生殖細胞-基礎から最前線まで(岡田益吉,長濱嘉孝編)" 共立出版(印刷中), (1995)

    • Related Report
      1994 Annual Research Report
  • [Publications] Pierce,D.F.: "Inhibition of mammary duct development but not alveolar outgrowth during pregnancy in MMTV-TGFβ^<S223/225> transgenic mice" Genes & Development. 7. 2308-2317 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] Coffey,R.J.: "Acceleration of mammary neoplasia in virgin mouse mammary tumor virus enhancer/promotor-transforming growth factor α transgenic mice by 7,12 dimethy benzanthracere" Cancer Research. (1994)

    • Related Report
      1993 Annual Research Report
  • [Publications] Yanai,N.: "Apoptosis induced at nonpermisive temperature by a transient increase in p53 in cell lines immortalized with ts SV40 T-antigen gene" Experimental Cell Research. 211. (1994)

    • Related Report
      1993 Annual Research Report
  • [Publications] Yanai,N.: "Role for integrin VLA-4 in stroma dependent erythnpoiesis" Blood. 83. (1994)

    • Related Report
      1993 Annual Research Report
  • [Publications] 松居靖久: "始原生殖細胞由来の細胞株の胚幹細胞としての性質" 日本疾患モデル学会記録. 9. 9-14 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] 松居靖久: "始原生殖細胞株の樹立とその性質" Tissue Culture Research Communications. 12. 273-278 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] 松居靖久: "生殖細胞の癌,テラトーマの発生機構" 実験医学. 11. 713-716 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] 松居靖久: "マウス始原生殖細胞の培養" 組織培養. 19. 176-180 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] 松居靖久: "Sl因子による始原生殖細胞の増殖制御機構" Medical Immunology. 26. 81-88 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] 松居靖久: "c-KitとSl因子の発生過程における発現と機能" 実験医学. 11. 1686-1691 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] 松居靖久: "c-Kitによる細胞内情報伝達" 生化学. 65. 1317-1321 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] 松居靖久: "マウス始原生殖細胞の体外培養" 実験医学. 12. 123-130 (1994)

    • Related Report
      1993 Annual Research Report
  • [Publications] 松居靖久: "生殖細胞の生成と増殖,分化" 細胞工学. (1994)

    • Related Report
      1993 Annual Research Report

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Published: 1993-04-01   Modified: 2016-04-21  

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