Journal of Embryo Transfer (한국수정란이식학회지)

The Korean Society of Embryo Transfer (한국수정란이식학회)
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Effect of Early Pregnant Heifer as Donor on the Ovum Pick-Up Derived Oocyte Aspiration and Embryo Production

초기 임신우의 공란우 활용이 초음파 유도 난자 채취 및 수정란 생산에 미치는 영향

  • Jin, Jong-In (Department of Animal Science, Division of Applied Life Science (BK21 Program), Graduate School of Gyeongsang National University) ;
  • Kwon, Tae-Hyeon (Department of Animal Science, Division of Applied Life Science (BK21 Program), Graduate School of Gyeongsang National University) ;
  • Choi, Byeong-Hyun (Department of Animal Science, Division of Applied Life Science (BK21 Program), Graduate School of Gyeongsang National University) ;
  • Kim, Sung-Soo (Department of Animal Science, Division of Applied Life Science (BK21 Program), Graduate School of Gyeongsang National University) ;
  • Jo, Hyun-Tea (Department of Animal Science, Division of Applied Life Science (BK21 Program), Graduate School of Gyeongsang National University) ;
  • Bang, Jae-Il (Department of Animal Science, Division of Applied Life Science (BK21 Program), Graduate School of Gyeongsang National University) ;
  • Kim, Sam-Chul (Department of Animal Science, Division of Applied Life Science (BK21 Program), Graduate School of Gyeongsang National University) ;
  • Cho, Kyu-Woan (College of Veterinary Medicine, Gyeongsang National University) ;
  • Lee, Jung-Gyu (Department of Animal Science, Division of Applied Life Science (BK21 Program), Graduate School of Gyeongsang National University) ;
  • Kong, Il-Keun (Department of Animal Science, Division of Applied Life Science (BK21 Program), Graduate School of Gyeongsang National University)
  • 진종인 (경상대학교 응용생명과학부 축산학(BK21)) ;
  • 권태현 (경상대학교 응용생명과학부 축산학(BK21)) ;
  • 최병현 (경상대학교 응용생명과학부 축산학(BK21)) ;
  • 김성수 (경상대학교 응용생명과학부 축산학(BK21)) ;
  • 조현태 (경상대학교 응용생명과학부 축산학(BK21)) ;
  • 방재일 (경상대학교 응용생명과학부 축산학(BK21)) ;
  • 김삼철 (경상대학교 응용생명과학부 축산학(BK21)) ;
  • 조규완 (경상대학교 수의과대학) ;
  • 이정규 (경상대학교 응용생명과학부 축산학(BK21)) ;
  • 공일근 (경상대학교 응용생명과학부 축산학(BK21))
  • Received : 2011.02.24
  • Accepted : 2011.03.02
  • Published : 2011.03.31
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Abstract

This study was carried out to evaluate the effect of early pregnant cow as donor for Ovum Pick-Up (OPU) derived oocyte aspiration and embryo production in Holstein heifers. Four non-pregnant and 2 pregnant Holstein heifers were used as donor and then carried out total 17 OPU session for 10 weeks (2 times per week). Recovered cumulus-oocyte-complexes (COCs) were classified into 4 grade by oocyte cytoplasm and cumulus cells and matured in vitro in TCM-199 supplemented with 10% FBS, 10 mg/ml FSH and 1 mg/ml estradiol in 5% $CO_2$ and over 99% humidity for 24 h. After 24 h co-incubation with post-thaw sperm, the presumed zygotes were cultured in CR1aa medium with 4 mg/ml BSA for 3 days and then changed CR1aa medium with 10% of FBS for another 3~4 days. The Mean number of aspirated follicles and collected oocytes in the early stage pregnant and non-pregnant heifers were $13.0{\pm}4.3$ and $10.6{\pm}3.9$, $5.4{\pm}3.4$ and $7.7{\pm}3.6$ per session, respectively. Rate of collected oocyte from aspirated follicles were 59.2% and 50.5%, respectively. The average number of good quality oocytes (Grade I and II) in the early stage pregnant and non-pregnant heifers was $3.7{\pm}2.7$ and $4.9{\pm}2.6$ (Mean${\pm}$SD). Cleavage and blastocyst developmental rates in Grade I and II were 22.2% and 25.5%, and then $1.7{\pm}0.9$ and $1.4{\pm}1.1$ blastocyst per session, respectively. In conclusion, OPU technology can be used in early stage pregnant and non-pregnant heifers without any problem and so applied OPU derived embryo production to maximize the ability of genetically valuable females.

본 연구에서는 유전적 가치가 높은 가축을 OPU 기술을 이용하여 단기간에 반복적이고 연속적으로 생체 내 난포란을 채란하여 수정란을 생산할 수 있는 가능성을 연구한 것으로써, 수정란의 안정적인 생산 및 이식으로 가축 개량의 세대 간격을 단축하기 위해 우수한 유전력을 가진 염선된 개체의 임신 초기 3개월 동안에 반복적인 수정란 생산 가능 여부를 조사하였다. 1. 비임신우 및 임신 초기우에 2회/주 채란으로 비인신우에서는 68회 채란으로 생성된 난포수는 721개($10.6{\pm}3.9$)의 난포를 흡입하여 364개($5.4{\pm}3.4$)의 난자를 회수하여 50.5%의 회수율을 보였으나, 임신 초기우에서는 34회 채란으로 441개($13.0{\pm}4.3$) 생성된 난포를 흡입하여 그중 261개($7.7{\pm}3.6$)의 난자가 회수되어 회수율이 59.2%로 유의적으로 높게 나타났다. 2. 초기임신우와 비임신우간의 회수된 난자의 등급은 비임신우 및 임신우에서 평균 $5.4{\pm}3.4$개 및 $7.7{\pm}3.6$개 회수된 난자에서 Grade II(평균 $2.1{\pm}2.1$개 및 $2.4{\pm}2.2$개)와 Grade IV(평균 $0.8{\pm}1.3$ 및 $1.2{\pm}1.5$개)는 유의적인 차이가 없었으나, Grade I(평균 $1.6{\pm}1.2$ 및 $2.4{\pm}1.3$개)과 Grade III(평균 $0.9{\pm}1.4$ 및 $1.6{\pm}1.9$개)은 유의차가 있었으며, Grade I, II 등급 출현율는 각각 $3.7{\pm}2.7$개 및 $4.9{\pm}2.6$개로 채취한 임선우에서 유의적으로 높게 나타났다. 3. 임신 초기우 및 비임신우로부터 채취한 난포란을 각 개체별 체외성숙 및 체외수정 후 배반포의 발생률은 수정 및 분할률에서 평균 $3.3{\pm}2.4$개, 임신우에서는 평균 $4.8{\pm}2.3$개로 유의적으로 높게 나타났으나, 이식가능한 배반포 발달은 93개 및 58개, 발달률은 25.5% 및 22.2%로 임신우에서 낮았으며, 또한 채란회수당 평균 배반포 발달에서는 $1.4{\pm}1.1$개 및 $1.7{\pm}0.9$개로 임신우의 평균 수가 많았으나 유의차는 없었다. 이상의 본 연구의 결과에서 가축의 임신 초기 개체에서도 1주일에 2회 반복적으로 OPU를 실시한 후 체외성숙 및 체외 배양을 통해 반복적으로 이식 가능한 수정란을 생산하였다. 이러한 방법은 가축의 초기 임신우로부터 채란이 가능하므로 개체의 임신에 대한 불안삼을 해소할 수 있으며, 유전적으로 아주 우수한 고능력우를 임신 기간 중에도 공란우로 이용하여 반복적으로 수정란의 생산이 가능하므로 그 이용의 활용도가 높을 것으로 사료된다.

Keywords

References

  1. Bols PEJ, Vandenheede JMM, van Soom A and de Kruif A. 1995. Transvaginal ovum pick-up (OPU) in the cow: A new disposable needle guidance system. Theriogenology 43:677-687. https://doi.org/10.1016/0093-691X(94)00073-4
  2. Brogliatti GM and Adams GP. 1996. Ultrasound-guided transvaginal oocyte collection in prepubertal calves. Theriogenology 45:1163-1176. https://doi.org/10.1016/0093-691X(96)00072-6
  3. Broussard JR, Rocha A, Lim JM, Blar RM, Roussel JD and Hansel W. 1996. The effect of environmental temperature and humidity on the quality and developmental competence of bovine oocytes obtained by transvaginal ultrasound-guided aspiration. Theriogenology 45:351 (Abstr). https://doi.org/10.1016/0093-691X(96)84824-2
  4. Bungartz L, Lucas-Hahn D, Rath D and Niemann H. 1995. Collection of oocytes from cattle via follicular aspiration aided by ultrasound with or without gonadotropin pretreatment and different reproductive stages. Theriogenology 13: 667-675.
  5. Chaubal SA, Molina JA, Ohlrichs CL, Ferre LB, Faber DC, Bols PEJ, Riesen JW, Tian X and Yang X. 2006. Comparison of different transvaginal ovum pick-up protocols to optimise oocyte retrieval and embryo production over a 10-week period in cows. Theriogenology 65:1631-1648. https://doi.org/10.1016/j.theriogenology.2005.07.020
  6. Duby RT, Damiani P, Looney CR, Fissore RA and Robl JM. 1996. Prepuberal calves as oocyte donors: Promises and problems. Theriogenology 45:121-130. https://doi.org/10.1016/0093-691X(95)00361-B
  7. Gibbons JR, Beal WE, Krisher RL, Faber EG, Pearson RE and Gwadauskas FC. 1994. Effects of once versus twice weekly transvaginal follicular aspiration on bovine oocyte recovery and embryo development. Theriogenology 41:206 (Abstr). https://doi.org/10.1016/S0093-691X(05)80116-5
  8. Gibbons JR, Krisher RL, Carlin RE, Pearson RE and Gwadauskas FC. 1995. In vitro embryo production after microinjection and ovarian dynamics following transvaginal follicular oocyte aspiration. Theriogenology 43:1129-1139. https://doi.org/10.1016/0093-691X(95)00076-K
  9. Guyader JC, Ponchon S, Thuard JM, Durand M, Nibart M, Marquant-Le Guiemme B and Hombolt P. 1997. Effects of superovulation on repeated ultrasound guided oocyte collection and in vitro embryo production in pregnant heifers. Theriogenology 47:157 (Abstr). https://doi.org/10.1016/S0093-691X(97)82284-4
  10. Hasler JF, Henderson WB, Hurtgen PJ, Mc-Cauley AD, Hower SA, Shuey LS, Stokes JE and Trimmer SA. 1995. Production, freezing and transfer of bovine IVF embryos and subsequent calving results. Theriogenology 43:141-159. https://doi.org/10.1016/0093-691X(94)00020-U
  11. Jeong WJ, Cho SJ, Lee HS, Deb GK, Lee YS, Kwon TH and Kong IK. 2009. Effect of cytoplasmic lipid content on in vitro developmental efficiency of bovine IVP embryos. Theriogenology 72:584-589. https://doi.org/10.1016/j.theriogenology.2009.04.015
  12. Kruip ThAM, Boni R, Roelofsen MWM, Wurth YA and Pieterse MC. 1994. Application of OPU for embryo production and breeding in cattle. Theriogenology 39:251 (Abstr).
  13. Lonergan P, Fair T, Corcoran D and Evans ACO. 2006. Effect of culture environment on gene expression and developmental characteristics in IVF-derived embryos. Theriogenology 65:137-152. https://doi.org/10.1016/j.theriogenology.2005.09.028
  14. Looney CR, Lindsey BR, Gonseth CL and Johnson DL. 1994. Commercial aspects of oocyte retrieval and in vitro fertilization (IVF) for embryo production in problem cow. Theriogenology 41:67-72. https://doi.org/10.1016/S0093-691X(05)80050-0
  15. Meintjes M, Bellow MS, Broussard JR, Paul JB and Godke RA. 1995. Transvaginal aspiration of oocytes from hormonetreated pregnant beef cattle for in vitro fertilization. J. Anim. Sci. 73:967-974. https://doi.org/10.2527/1995.734967x
  16. Merton JS, de Roos AP, Mullaart E, de Ruigh L, Kaal L, Vos PL and Dieleman SJ. 2003. Factors affecting oocyte quality and quantity in commercial application of embryo technologies in the cattle breeding industry. Theriogenology 59:651-674. https://doi.org/10.1016/S0093-691X(02)01246-3
  17. Petyim S, Bage R, Hallap T, Bergqvist AS, Rodriguez-Martinez H and Larsson B. 2003. Two different schemes of twiceweekly ovum pick-up in dairy heifers: Effect on oocyte recovery and ovarian function. Theriogenology 60:175-188. https://doi.org/10.1016/S0093-691X(02)01363-8
  18. Pieterse MC, Kappen KA, Kruip TM and Taverne MA. 1988. Aspiration of bovine oocytes during transvaginal ultrasound scanning of the ovaries. Theriogenology 30:751-762. https://doi.org/10.1016/0093-691X(88)90310-X
  19. Pieterse MC, Vos PLAM, Kruip TM, Willemse AH and Taverne MA. 1991a. Characteristics of bovine estrrus cycles during repeated transvginal, ultrasound-guided puncturing of follicles for ovum pick-up. Theriogenology 35:401-413. https://doi.org/10.1016/0093-691X(91)90290-T
  20. Pieterse MC, Vos PLAM, Kruip TM, Wruth YA, van Beneden ThH, Willemse AH and Taverne MAM. 1991b. Transvaginal ultrasound guided follicular aspiration bovine oocytes. Theriogenology 35:19-24. https://doi.org/10.1016/0093-691X(91)90144-3
  21. Pieterse MC, Vos PLAM, Kruip TM, Wurth YA, van Beneden TH, Willemse AH and Taverne MA. 1992. Repeated transvaginal ultrasound-guided ovum pick-up in eCG-treated cow. Theriogenology 37:273 (Abstr). https://doi.org/10.1016/0093-691X(92)90342-O
  22. Presicce GA, Jiang, S, Simken M, Zhang L, Looney CR, Godke RA and Yang X. 1997. Age and hormonal dependence of acquisition of oocyte competence for embryogenesis in prepubertal calves. Biol. Reprod. 56:386-392. https://doi.org/10.1095/biolreprod56.2.386
  23. Reinders JMC and van Wagtendonk-de Leeuw AM. 1996. Improvement of a moet program by addition of in vitro production of embryos after ovum pick up from pregnant donor heifers. Theriogenology 45:354 (Abstr). https://doi.org/10.1016/0093-691X(96)84827-8
  24. Scott CA, Robertson L, Mousra RTD, Paterson C and Boyd JS. 1994. Technical aspects of transvaginal ultrasound-guided follicular aspiration in cows. Vet. Rec. 134:440-443. https://doi.org/10.1136/vr.134.17.440
  25. Stubbings RB and Walton JS. 1995. Effect of ultrasonicallyguided follicle aspiration on estrous cycle and follicular dynamics in Holstein cows. Theriognology 43:705-712. https://doi.org/10.1016/0093-691X(95)00013-X
  26. Thibier M. 2008. The worldwide activity in farm animals embryo transfer. Embryo Transfer Newsletter (IETS) 26:4-9.
  27. Van Wagtendonk-de Leeuw AM, Mullaart E, Roos de APW, Merton JS, Den Daas JHG and de Ruigh L. 2000. Effects of different reproduction techniques: AI, MOET or IVP, on health and welfare of bovine offspring. Theriogenolgy 53:575-597. https://doi.org/10.1016/S0093-691X(99)00259-9
  28. Walton JS, Christie KA and Stubbings RB. 1993. Evaluation of frequency of ultrasonically guided follicle aspiration on ovarian dynamics. Theriognology 39:336 (Abstr). https://doi.org/10.1016/0093-691X(93)90191-7
  29. Willett EL, Black WG, Casida LE, Stone WH and Buckner PJ. 1951. Successful transplantation of a fertilized bovine ovum. Science 113:247.
  30. 이병천, 윤기영, 김정태, 이강남, 노상호, 신태영, 박종임, 김남렬, 주석천, 백남용, 이은송, 임정묵, 이우근, 황우석, 1998. 초음파 유도에 의한 소 난포란의 채취에 관한 연구. 2. 임신우 유래 난포란으로 부터 산자생산에 관하여. 한국수정란학회지 13:77-86.