Journal of Embryo Transfer (한국수정란이식학회지)

The Korean Society of Embryo Transfer (한국수정란이식학회)
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Production of Korean Native Cow from Mongolian Cow following Transfer of Vitrified Blastocyst

Mongolian 수란우에 한우 동결수정란의 이식 후 산자 생산

  • Kong, I.K. (Department of Animal Science and Technology, Sunchon National University) ;
  • Sanjjav, G. (Department of Physiology, Monogolia State University of Agriculture) ;
  • Yang, C.J. (Department of Animal Science and Technology, Sunchon National University) ;
  • Cho, S.G. (Department of Animal Science and Technology, Sunchon National University) ;
  • Bae, I.H. (Department of Animal Science and Technology, Sunchon National University) ;
  • Oh, D.H. (Department of Animal Science and Technology, Sunchon National University)
  • Published : 2002.08.01
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Abstract

The purpose of this study was to investigate the comparison of viability of bovine blastocysts following glass micropipette (GMP) vitrification and the possibility of production of Korean Native Cow ("Hanwoo,"Bos taurus coreanae) following embryo transfer into Mongolia cows (Bos taurus mongolian). The embryos of Korean Native Cow were produced by IVMFC or superovulation methods in Korea, cryopreserved by GMP vitrification, and subsequently trans-ported to Mongolia. The recipient cows were synchronized using a CIDR plus and prostaglandin $F_2\alpha$($PGF_2\alpha$) treatment. To produce in vivo embryos, seven cows were superovulated using FSH and PGF$_2$／sub $\alpha$／ treatment. A total of 64 blastocysts ( $9.1\pm2.94$ per cow) were collected. In vitro embryos were produced using a defined culture system which cleaved in 80.1％ ova (174／217), and developed to blastocyst stage embryos of 40.8％ (71／174). The post-thaw survival rate of in vivo blastocysts (93.7％; 45／48) was significantly higher than that of in vitro blastocysts (82.5％; 52／63, P＜0.05). Embryo transfer was carried out using 8 Mongolian recipient cows and 2 post-thaw blastocysts per recipient. Five of 8 recipients were found pregnant at Day 60 but one abortion occurred by Day 240. Two of offspring were produced from the Mongolian cows at 275 days after embryo transfer. These results indicated that a GMP vitrification method could be used as a cryopreservation technique for in vivo or in vitro bovine blastocysts and produced effectively a Korean Native Cow following embryo transfer into a Mongolian recipient cow.

본 연구는 체내, 체외 소 배반포기 배의 GMP vitrification 후 활력도의 비교와 한우 수정란을 몽골 소에 수정란이식 후 산자생산 가능성을 조사하고자 실시하였다. 한우 수정란은 체외수정란 또는 과배란처리에 의한 체내수정란을 생산하여 GMP vitrification 방법으로 동결 후 몽고로 수송하였다. 수란우는 CIDR과 $PGF_2\alpha$ 처리에 의하여 동기화를 유도하였다 체내수정란생산을 위하여 7두를 과배란처리하였다. 총 64개의 배반포기를 회수하였다. ($9.1\pm2.94$per cow). 체외수정란생산은 80.1% 분할율(174／217)과 40.8% 배반포기 발달율(71／174)을 얻었다. 체내수정란(93.7%; 45／48)의 동결융해 후 생존율은 체외수정란(82.5%; 52／63)보다 유의적으로 높았다(P＜0.05). 8두의 몽골 소에 2개의 수정란을 이식하여 5두가 이식 후 60일째 임신이 확인되었으나, 그 중 1두는 240째 유산을 확인하였다. 그 중 2두의 수란우에서 2두의 산자를 275일째 생산에 성공하였다. 이러한 결과는 GMP vitrification 방법은 체내, 체외수정란의 동결보존방법으로 이용될 수 있을 뿐만 아니라 동결융해란의 몽골 소에 이식 후 한우를 생산할 수 가능성을 확인하였다.

Keywords

References

  1. Agca Y, Monson RL, Northey DL, Mazni AO and Rutledge JJ. 1994. Post-thaw survival and pregnancy rates of in vitro produced bovine embryos after vitrification. Theriogenology, 41:154(Abstr.) https://doi.org/10.1016/S0093-691X(05)80064-0
  2. Agca Y, Monson RL, Northey DL, Mazni AO, Schaefer DM and Rutledge JJ. 1998. Transfer of fresh and cryopreserved IVP bovine embryos: normal calves, birth weight and gestation lengths. Theriogenology, 50:147-162 https://doi.org/10.1016/S0093-691X(98)00121-6
  3. Fissore RA, Dobrinsky JR, Balise JJ, Duby RT and Robl JM. 1992. Patterns of intracellular $Ca^{2+}$ concentration in fertilized bovine eggs. Biol. Reprod., 47:960-969 https://doi.org/10.1095/biolreprod47.6.960
  4. Greve T, Avery B and Callesen H. 1993. Viability of in vivo and in vitro produced bovine embryos. Reprod. Domest. Anim., 28:164-169 https://doi.org/10.1111/j.1439-0531.1993.tb00113.x
  5. Jackowski SC, Leibo SP and Mazur P. 1980. Glycerol permeability of fertilized and unfertilized mouse ova. J. Exp. Zool., 212:329-341 https://doi.org/10.1002/jez.1402120305
  6. Kong IK, Lee SI, Cho SG, Cho SK and Park CS. 2000. Comparison of open pulled straw (OPS) vs glass micropipette (GMP) vitrification in mouse blastocysts. Theriogenology, 53:1817-1826 https://doi.org/10.1016/S0093-691X(00)00317-4
  7. Leibo SP. 1980. Water permeability and its activation energy of fertilized and unfertilized mouse ova. J. Membr. Biol., 53:179-188 https://doi.org/10.1007/BF01868823
  8. Leibo SP and Loskutoff NM. 1993. Cryobiology of in vitro derived bovine embryo. Theriogenology, 39:81-94 https://doi.org/10.1016/0093-691X(93)90025-Z
  9. Leibo SP, Martino A, Kobayashi S and Pollard JW. 1996. Stage dependent sensitivity of oocytes and embryos to low temperatures. Anim. Reprod. Sci., 42:45-53 https://doi.org/10.1016/0378-4320(96)01543-6
  10. Machaty Z, Day BN and Prather RS. 1998. Development of early porcine embryos in vitro and in vivo. Biol. Reprod., 59:451-455 https://doi.org/10.1095/biolreprod59.2.451
  11. Mahmoudzadeh AR, Van Soom A, Bols P, Ysebaert MT and Kruif A. 1995. Optimization of a single vitrification procedure for bovine embryos produced in vitro: effect of developmental stage, two step addition of cryoprotectant and sucrose dilution on embryonic survival. J. Reprod. Fertil., 103:33-39 https://doi.org/10.1530/jrf.0.1030033
  12. Massip A, Mermillod P and Dinnyes A. 1995a. Morphology and biochemistry of in vitro procedure bovine embryos: implication for their cryopreservation. Hum. Reprod., 10:3004-3011 https://doi.org/10.1093/oxfordjournals.humrep.a135837
  13. Massip A, Mermillod P, Van Langendonckt A, Touze JL and Dessy F. 1995b. Survival and viability of fresh and frozen-thawed in vitro bovine blastocysts. Reprod. Nutr. Dev., 35:3-10 https://doi.org/10.1051/rnd:19950101
  14. Massip A, van der Zwalmen P, Scheffen B and Ectors F. 1986. Pregnancies following transfer of cattle embryos preserved by vitrification. Cryo-Letters, 7:270-273
  15. McKieman SH, Clayton MK and. Bavister BD. 1995. Analysis of stimulatory and inhibitory amino acids for development of hamster one-cell embryos in vitro. Mol. Reprod. Dev., 42:188-199 https://doi.org/10.1002/mrd.1080420208
  16. Niemann H. 1991. Cryopreservation of ova and embryos from livestock: current status and research needs. Theriogenology, 35:109-124 https://doi.org/10.1016/0093-691X(91)90151-3
  17. Parrish JJ, Susko-Parrish JL, Wimer MA and First NL. 1988. Capacitation of bovine sperm by heparin. Biol. Reprod., 38:1171-1180 https://doi.org/10.1095/biolreprod38.5.1171
  18. Pollard JW and Leibo SP. 1993. Comparative cryobiology of in vitro and in vivo derived bovine embryos. Theriogenology, 39:287 (Abstr.) https://doi.org/10.1016/0093-691X(93)90142-R
  19. Rall WF and Fahy GM. 1985. Ice-free cryopreservation of mouse embryos at -196${^\circ}C$ by vitrification. Nature, 313:573-575 https://doi.org/10.1038/313573a0
  20. SAS Institute Inc. 1990. The GLM procedure. SAS/STAT User's Guide. Version 6 (4th Edn, Vol. 2), Cary, NC, USA, SAS Institute Inc., pp. 891-996
  21. Tervit HR, Pugh PA, McGowan LT, Bell ACS and Wells RW. 1994. The freezability of sheep embryos is affected culture system and source (in vivo and in vitro derived). Theriogenology, 41:315 (Abstr.) https://doi.org/10.1016/S0093-691X(05)80225-0
  22. Tsuzuki Y, Go J, Fujihara N and Ashizawa K. 1992. Ultrastructural studies of bovine blastocysts fertilized in vitro associated with freezing and thawing. Proc. 12th Inter. Cong, Anim. Reprod., 3:1487-1489
  23. Vajta G, Holm P, Greve T and Callesen H. 1996. Overall efficiency of in vitro embryo production and vitrification in cattle. Theriogenology, 45:683-689 https://doi.org/10.1016/0093-691X(95)00414-4
  24. Vajta G, Holm P, Greve T and Callesen H. 1997. Survival and development of bovine blastocysts produced in vitro after assisted hatching, vitrification and in straw direct rehydration. J. Reprod. Fertil., 111:65-70 https://doi.org/10.1530/jrf.0.1110065
  25. Vajta G, Holm P, Kuwayama M, Booth PJ, Jacobsen H, Greve T and Callesen H. 1998. Open pulled straw (OPS) vitrification: a new way to reduce cryoinjuries of bovine ova and embryos. Mol. Reprod. Dev., 51:53-58 https://doi.org/10.1002/(SICI)1098-2795(199809)51:1<53::AID-MRD6>3.0.CO;2-V