JARB Journal of Animal Reproduction and Biotehnology

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eISSN: 2671-4663

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Journal of Embryo Transfer 2008; 23(3): 223-227

Published online September 30, 2008

Copyright © The Korean Society of Animal Reproduction and Biotechnology.

한우 수정란의 동결보존 후 발달 효율 비교

조상래, 최창용, 김현종, 최선호, 손동수

농촌진흥청 축산과학원 가축유전자원시험장

Abstract

The cryopreservation of Hanwoo embryos has become an integral part of assisted reproduction in animal. The objective of this study was to assess the effect of The objectives of this study were: (1) to evaluate the influence of bovine embryo developmental stage on in vitro embryo development after freezing, (2) to study the efficiency compared with conventional freezed embryos at different embryo source. For conventional slow-freezing, day 7 or 8 expanded blastocysts were collected. The standard freezing medium was 1.8 M ethylene glycol (EG). Embryos were equilibrated in 1.8 Methylene glycol(EG) with 0.1 M sucrose in Dulbecco's phosphate-buffered saline (D-PBS) supplemented with 0.5% bovine serum albumin. Embryos were then loaded individually into 0.25 ml-straw and placed directly into cooling chamber of programmable freezer precooled to $-7^{circ}C$, after 2 min, the straw was seeded, maintained at $-7^{circ}C$ for 8 min, and then cooled to $-35^{circ}C$ at $0.3^{circ}C$/min, plunged and stored in liquid nitrogen for at least 3 days. For thawing, the straw containing embryos were warmed in air for 10 see and exposed to $37^{circ}C$ water for 20 sec. Straws were then removed from $37^{circ}C$ water. Rates of blastocyst survive and hatched were evaluated at 12 to 48h post-warming. The re-expansion and hatched rates of morula embryos were significantly lower than those obtained for blastocysts and expansion blastocysts (31.6%, 10.5% vs, 68.9%, 22.2% vs, 73.7%, 53.6%, respectively). No differences in re-expansion rates were found between in vivo and in vitro blastocysts. whereas hatched rates was significantly higher (51.2%) in vivo compared with in vitro embryos (18.6%). in conclusion, demonstrate that conventional freezing can be used successfully in cryopreservation of in vitro and in vivo bovine embryos, and that it might be considered for use in commercial programs and embryo preservation.

Keywords: bovine, embryos, in vitro, in vivo, cryopreservation

Article

Journal of Embryo Transfer 2008; 23(3): 223-227

Published online September 30, 2008

Copyright © The Korean Society of Animal Reproduction and Biotechnology.

한우 수정란의 동결보존 후 발달 효율 비교

조상래, 최창용, 김현종, 최선호, 손동수

농촌진흥청 축산과학원 가축유전자원시험장, 농촌진흥청 축산과학원 가축유전자원시험장, 농촌진흥청 축산과학원 가축유전자원시험장, 농촌진흥청 축산과학원 가축유전자원시험장, 농촌진흥청 축산과학원 가축유전자원시험장

한우 수정란의 동결보존 후 발달 효율 비교

조상래, 최창용, 김현종, 최선호, 손동수

농촌진흥청 축산과학원 가축유전자원시험장

Abstract

The cryopreservation of Hanwoo embryos has become an integral part of assisted reproduction in animal. The objective of this study was to assess the effect of The objectives of this study were: (1) to evaluate the influence of bovine embryo developmental stage on in vitro embryo development after freezing, (2) to study the efficiency compared with conventional freezed embryos at different embryo source. For conventional slow-freezing, day 7 or 8 expanded blastocysts were collected. The standard freezing medium was 1.8 M ethylene glycol (EG). Embryos were equilibrated in 1.8 Methylene glycol(EG) with 0.1 M sucrose in Dulbecco's phosphate-buffered saline (D-PBS) supplemented with 0.5% bovine serum albumin. Embryos were then loaded individually into 0.25 ml-straw and placed directly into cooling chamber of programmable freezer precooled to $-7^{circ}C$, after 2 min, the straw was seeded, maintained at $-7^{circ}C$ for 8 min, and then cooled to $-35^{circ}C$ at $0.3^{circ}C$/min, plunged and stored in liquid nitrogen for at least 3 days. For thawing, the straw containing embryos were warmed in air for 10 see and exposed to $37^{circ}C$ water for 20 sec. Straws were then removed from $37^{circ}C$ water. Rates of blastocyst survive and hatched were evaluated at 12 to 48h post-warming. The re-expansion and hatched rates of morula embryos were significantly lower than those obtained for blastocysts and expansion blastocysts (31.6%, 10.5% vs, 68.9%, 22.2% vs, 73.7%, 53.6%, respectively). No differences in re-expansion rates were found between in vivo and in vitro blastocysts. whereas hatched rates was significantly higher (51.2%) in vivo compared with in vitro embryos (18.6%). in conclusion, demonstrate that conventional freezing can be used successfully in cryopreservation of in vitro and in vivo bovine embryos, and that it might be considered for use in commercial programs and embryo preservation.

Keywords: bovine, embryos, in vitro, in vivo, cryopreservation

JARB Journal of Animal Reproduction and Biotehnology

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OPEN ACCESS pISSN: 2671-4639
eISSN: 2671-4663