Survival of Porcine Delipated Oocytes and Embryos after Cryopreservation by Freezing or Vitrification

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概要

• 論文の詳細を見る

• The present study examined whether delipated porcine oocytes and embryos at various stages of development can be cryopreserved by conventional slow cooling or vitrification. Most (93%) of the 27 delipated morulae developed to blastocysts after freezing with 1.5 M propanediol + 0.1 M sucrose. Late morulae and early blastocysts delipated at 2-4 cell stage and cultured in vitro survived freezing either with 1.5 M glycerol + 0.25 M sucrose (10/18, 56%) or 1.8 M ethylene glycol + 0.25 M sucrose (14/19, 74%). Delipated 2-4 cell stage embryos and oocytes could be cryopreserved by vitrification with 40% ethylene glycol, 1 M sucrose and 20% fetal calf serum. Half (7/14) of the vitrified, delipated embryos developed to blastocysts after thawing. Of 48 delipated oocytes, 27 (56%) maintained an intact outline of the ooplasm after vitrification and underwent subzonal sperm injection. Fertilization was confirmed in 12 (25%) of these oocytes and 3 (6%) developed to morula stage. This study also aimed at developing a non-invasive method for cryopreserving porcine embryos after reducing their cytoplasmic lipid content without micromanipulation. Morulae and early blastocysts were centrifuged in the presence of cytochalasin B and cryoprotectants and then frozen immediately. More than half (14/24, 58%) of the centrifuged morulae developed to blastocycts after freezing with 1.5 M propanediol + 0.1 M sucrose. Greater than 70% of centrifuged early blastocysts survived freezing either with 1.5 M propanediol (30/31, 97%), 1.5 M glycerol (22/29, 76%) or 1.8 M ethylene glycol (21/29, 72%). These results demonstrated that delipation (lipid removal) from porcine oocytes and embryos at various stages enables their cryopreservation. A new insight into the development of a non-invasive method for cryopreserving porcine embryos was also provided.

• 日本繁殖生物学会の論文

著者

• Kuwayama M

  Kato Ladies Clinic Tokyo Jpn

• Kuwayama Masashige

  Animal Biotechnology Center Livestock Improvement Association Of Japan

• Nagashima Hiroshi

  BresaGen Limited, PO Box 259, Rundle Mall, Adelaide, S.A. 5000, Australia

• Cameron Renald

  Department of Farm Animal Medicine and Production, The University of Queensland, Queensland 4067, Au

• Young Mary

  Department of Farm Animal Medicine and Production, The University of Queensland, Queensland 4067, Au

• Beebe Luke

  Department of Farm Animal Medicine and Production, The University of Queensland, Queensland 4067, Au

• W. Blackshaw

  Department of Farm Animal Medicine and Production, The University of Queensland, Queensland 4067, Au

• B. Nottle

  BresaGen Limited, PO Box 259, Rundle Mall, Adelaide, S.A. 5000, Australia

• Nagashima H

  Meiji Univ. Kanagawa

• D.A. Cameron

  Department of Farm Animal Medicine and Production, The University of Queensland, Queensland 4067, Au

関連論文

• Survival of Porcine Delipated Oocytes and Embryos after Cryopreservation by Freezing or Vitrification
• Vitrification of bovine blastocysts derived from in vitro maturation, in vitro fertilization and in vitro culture
• Improved Survival of Porcine Hatched Blastocysts Cryopreserved with Glycerol and Sucrose
• Survival of Porcine Delipated Oocytes and Embryos after Cryopreservation by Freezing or Vitrification
• Improved Survival of Porcine Hatched Blastocysts Cryopreserved with Glycerol and Sucrose
• Effect of β-Merchaptoethanol on the Preimplantation Development of Bovine Embryos Fertilized In Vitro
• Tumorigenesis and Gene Transmission in Transgenic Mice Carrying MMTV/v-Ha-ras

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