JARB Journal of Animal Reproduction and Biotehnology

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

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Journal of Embryo Transfer 2013; 28(3): 169-175

Published online September 30, 2013

Copyright © The Korean Society of Animal Reproduction and Biotechnology.

Production of Transgenic Cattle by Non-surgical Embryo Transfer

엄상준, 양정석, 이수민, 조소영, 허영태, 허영남, 구본철, 정기수, 김광재, 김지태, 김남형, 고대환

상지영서대학교 동물생명산업과, 충북대학교 축산학과, 대구가톨릭대학교 의학과, 강원도가축위생시험소남부지소

Abstract

Recently, the transgenic animal production technique is very important for the production of bio-parmaceutical as animal bio-reactor system. However, the absence of survival evaluation in vitro produced transgenic embryos has been a problem of the low productivity of transgenic animal because of absent of pre-estimate of pregnancy after transgenic embryos transferred into recipient. Therefore, this study is conducted to improve efficiency of transgenic cattle production by improving the non-surgical embryo transfer (ET) method. Transgenic bovine embryos were produced by injection of feline immunodeficiency virus enhanced green fluorescent protein (FIV-EGFP) lentiviral vector into perivitelline space of in vitro matured MII stage oocytes, and then in vitro fertilization (IVF) was occured. Normal IVF and EGFP expressing blastocysts were transferred into recipients. Results indicated that 2 expanded blastocysts (34.7%) transferred group showed significantly (P<0.05) higher pregnancy rate than 1 expanded blastocyst (26.8%) transferred group. In case of parity of recipient, ET to heifer (34.9%) showed significantly (P<0.05) higher pregnancy rate than ET to multiparous recipient (21.2%). However, there are no significant differences of pregnancy rate between natural induced estrus and artificial induced estrus groups. Significantly (P<0.05) higher pregnancy rate was obtained from recipient group which have normal corpus luteum with crown group (34.8%) than normal corpus luteum without crown (13.6%). Additionally, treatment of $100{mu}g$ Gn-RH injection to recipient group (38.6%) 1 day before ET significantly (P<0.05) increase pregnancy rate than non- Gn-RH injection to recipient group (38.6%). We also transferred 2 EGFP expressing expanded blastocysts to each 19 recipients, 7 recipients were pregnant and finally 5 EGFP transgenic cattle were produced under described ET condition. Therefore, our result suggested that transfer of 2 good-quality expanded blastocysts to $100{mu}g$ of Gn-RH injected recipient which have normal corpus luteum with crown is feasible to produce transgenic cattle.

Keywords: embryo transfer, expand blastocyst, pregnancy, EGFP, transgenic cattle

Article

Journal of Embryo Transfer 2013; 28(3): 169-175

Published online September 30, 2013

Copyright © The Korean Society of Animal Reproduction and Biotechnology.

비외과적 수정란 이식에 의한 형질전환 소 생산 기술

엄상준, 양정석, 이수민, 조소영, 허영태, 허영남, 구본철, 정기수, 김광재, 김지태, 김남형, 고대환

상지영서대학교 동물생명산업과, 상지영서대학교 동물생명산업과, 상지영서대학교 동물생명산업과, 상지영서대학교 동물생명산업과, 충북대학교 축산학과, 충북대학교 축산학과, 대구가톨릭대학교 의학과, 강원도가축위생시험소남부지소, 강원도가축위생시험소남부지소, 강원도가축위생시험소남부지소, 충북대학교 축산학과, 상지영서대학교 동물생명산업과

Production of Transgenic Cattle by Non-surgical Embryo Transfer

엄상준, 양정석, 이수민, 조소영, 허영태, 허영남, 구본철, 정기수, 김광재, 김지태, 김남형, 고대환

상지영서대학교 동물생명산업과, 충북대학교 축산학과, 대구가톨릭대학교 의학과, 강원도가축위생시험소남부지소

Abstract

Recently, the transgenic animal production technique is very important for the production of bio-parmaceutical as animal bio-reactor system. However, the absence of survival evaluation in vitro produced transgenic embryos has been a problem of the low productivity of transgenic animal because of absent of pre-estimate of pregnancy after transgenic embryos transferred into recipient. Therefore, this study is conducted to improve efficiency of transgenic cattle production by improving the non-surgical embryo transfer (ET) method. Transgenic bovine embryos were produced by injection of feline immunodeficiency virus enhanced green fluorescent protein (FIV-EGFP) lentiviral vector into perivitelline space of in vitro matured MII stage oocytes, and then in vitro fertilization (IVF) was occured. Normal IVF and EGFP expressing blastocysts were transferred into recipients. Results indicated that 2 expanded blastocysts (34.7%) transferred group showed significantly (P<0.05) higher pregnancy rate than 1 expanded blastocyst (26.8%) transferred group. In case of parity of recipient, ET to heifer (34.9%) showed significantly (P<0.05) higher pregnancy rate than ET to multiparous recipient (21.2%). However, there are no significant differences of pregnancy rate between natural induced estrus and artificial induced estrus groups. Significantly (P<0.05) higher pregnancy rate was obtained from recipient group which have normal corpus luteum with crown group (34.8%) than normal corpus luteum without crown (13.6%). Additionally, treatment of $100{mu}g$ Gn-RH injection to recipient group (38.6%) 1 day before ET significantly (P<0.05) increase pregnancy rate than non- Gn-RH injection to recipient group (38.6%). We also transferred 2 EGFP expressing expanded blastocysts to each 19 recipients, 7 recipients were pregnant and finally 5 EGFP transgenic cattle were produced under described ET condition. Therefore, our result suggested that transfer of 2 good-quality expanded blastocysts to $100{mu}g$ of Gn-RH injected recipient which have normal corpus luteum with crown is feasible to produce transgenic cattle.

Keywords: embryo transfer, expand blastocyst, pregnancy, EGFP, transgenic cattle