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Serum-free cryopreservation of bovine mammary fragments-Effects of growth factors

Research Project

Project/Area Number 03660272
Research Category

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

Allocation TypeSingle-year Grants
Research Field 畜産学(含草地学)
Research InstitutionObihiro University of Agriculture and Veterinary Medicine

Principal Investigator

FURUMURA Keiko  Obihiro University of Agriculture and Veterinary Medicine Department of Animal Production and Agricultural Economics Instructor, 畜産学部, 助手 (50173547)

Project Period (FY) 1991 – 1992
Project Status Completed (Fiscal Year 1992)
Budget Amount *help
¥1,900,000 (Direct Cost: ¥1,900,000)
Fiscal Year 1992: ¥400,000 (Direct Cost: ¥400,000)
Fiscal Year 1991: ¥1,500,000 (Direct Cost: ¥1,500,000)
KeywordsSerum-free cryopreservation / bovine mammary fragments / Collagen gel culture / Insulin-like growth factor-I / Epidermal growth factor / Methylcellulose / 3H-thymidine incorporation / Autoradiography / 乳曽細胞増殖 / オートラジオグラフイ / コラ-ゲン・ゲル内包埋培養 / 凍結保存法 / 牛乳腺細胞増殖 / ^3Hーチミジン取り込み量 / メチルセルロ-ス / 牛胎子血清 / IGFーI / EGF
Research Abstract

1. It is considered that milk yield is depend on the number of mammary epithelial cells. However, the mechanism of proliferation and differentiation on bovine mammary epithelial cell has not been clarified. There is an economical limitation for using a cow as an experimental animal. In the slaughter house, bovine udders are usually thrown into the discard, therefore, we are able to get a lot of bovine mammary cells for these udders. It is beneficial that we cryopreserve the bovine mammary cells when we get a lot of fresh samples. Only a few reports have appeared about the culture of cryopreserved bovine mammary cells. All these reports have used serum in cryopreservation medium, and there is a possibility of undefined effect from serum to mammary cells.
The objectives were to: 1) evaluate the proliferative response of mammary fragments to serum-free cryopreservation medium, methylcellulose-demethylsulfoxide-medium 199 (MC-DMSO-M 199), 2) compare with fresh and cryopreserved mammary grow … More th response to synthetic insulinlike growth factor-1 (IGF-1) or epidermal growth factor (EGF) in vitro. Tritium (^3H) thymidine incorporation and morphological aspects were monitored to determine the proliferation of mammary cells in embedded collagen gel culture.
2.1) Pregnant 6 mice, lactating Holstein 3 cows, 6 steers and hormone treated (E/P) 4 ewes were used. 2) Mammary gland was digested with collagenase/hyaluronidase and mammary fragments were collected. One portion of fragments was used for fresh culture and other remaining fragments were frozen in serum-free or serum added cryopreservation medium. 3) After 6 and 12 months' cryopreservation, mammary cells were thawed immediately in 37C water and cultured in collagen gel.
3.1) Pregnant mouse mammary fragments grew well in collagen gel after 5 months serum-free cryopreservation and there is no growth difference between 0.1%MC cryopreserved cells and 0.05% MC cryopreserved cells. Therefore, 0.1% concentration of MC is used. 2) In fresh culture, lactating mammary fragments did not proliferated and ^3H-TdR incorporation was also very low. On the other hand, steer and E/P treated ewe mammary fragments grew rapidly and they also showed high ^3H-TdR incorporation response to IGF-1 and EGF. 3) After 6 and 12 months' serum-free cryopreservation, steer and E/P ewe mammary fragments proliferated well and showed the same morphological change as the one of fresh culture. ^3H-TdR after cryopreservation also showed the high response to the growth factors. Less

Report

(3 results)
  • 1992 Annual Research Report   Final Research Report Summary
  • 1991 Annual Research Report
  • Research Products

    (13 results)

All Other

All Publications (13 results)

  • [Publications] 古村 圭子,大山 勝彦,新出 陽三: "培養乳腺細胞の増殖に及ぼす成長因子(EGF及びIGF-I)添加の効果" 日本畜産学会第85回大会講演要旨. 3-18 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] 古村 圭子,澤田 妃佐江,新出 陽三: "無血清凍結保存後のコラーゲン・ゲル内包埋培養乳腺細胞の増殖" 日本畜産学会第87回大会講演要旨. V11-33 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] 古村 圭子: "コラーゲン・ゲル内包埋培養における牛乳腺細胞の増殖" 栄養生理研究会報. 37. 87-114 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] K.FURUMURA,H.SAWADA & Y.SHINDE: "Mammary cell growth after serum-free cryopreservation in collagen gel culture." VII World COnference on Animal Production,June 28-July 2. (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Keiko FURUMURA, Katsuhiko OHYAMA and Yozo SHINDE: "Effects of growth factors (EGF and IGF-I) on proliferation of mammary fragments in collagen gel culture." Japanese Society of Zootechnical Science, The 85 Annual Meeting Abstracts. (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Keiko FURUMURA, Hisae SAWADA and Yozo SHINDE: "Proliferation of Mammary fragments after serum-free cryopreservation in collagen gel culture." Japanese Society of Zootechnical Science, The 87 Annual Meeting Abstracts. (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Keiko FURUMURA: "In vitro proliferation of bovine mammary fragments in collagen gel culture." Proceedings of Japanese for Animal Nutrition and Metabolism. 37(1). 87-114 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Keiko FURUMURA, Hisae SAWADA and Yozo SHINDE: "Mammary cell growth after serum-free cryopreservation in collagen gel culture." VII World Conference on Animal Production, June 28-July 2. (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] 古村 圭子・大山 勝彦・新出 陽三: "培養乳腺細胞の増殖に及ぼす成長因子(EGF及びIGF-I)添加の効果" 日本畜産学会第85回大会講演要旨. P3-18 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] 古村 圭子・澤田 妃佐江・新出 陽三: "無血清凍結保存後のコラーゲン・ゲル内包埋培養乳腺細胞の増殖" 日本畜生学第87回大会講演要旨. V11-33 (1993)

    • Related Report
      1992 Annual Research Report
  • [Publications] 古村 圭子: "コラーゲン・ゲル内包埋培養における牛乳腺細胞の増殖" 栄養生理研究会報. 37(1). 87-114 (1993)

    • Related Report
      1992 Annual Research Report
  • [Publications] K.Furumura. H.Sawada & Y.Shinde: "Mammary cell growth after serum-free cryopreservation in collagen gel culture." 7 World Conference on Animal Production,June 28-July 2. (1993)

    • Related Report
      1992 Annual Research Report
  • [Publications] 古村 圭子,大山 勝彦,新出 陽三: "培養乳腺細胞の増殖に及ぼす成長因子(EGF及びIGFーI)添加の効果" 日本畜産学会第85回大会講演要旨. P3-18 (1992)

    • Related Report
      1991 Annual Research Report

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Published: 1991-04-01   Modified: 2016-04-21  

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