Journal of Embryo Transfer (한국수정란이식학회지)

The Korean Society of Embryo Transfer (한국수정란이식학회)
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Effects of Some Factors on In Vitro Production of Embryos from Antral Follicle-Derived Porcine Oocytes IV. Effects of Development Media and Those Change on In Vitro Development

돼지 난포란 유래 체외수정란 생산에 대한 제요인의 영향 IV. 체외발달 배양액의 종류와 배양액 교체가 체외발달에 미치는 영향

  • 연성흠 (농촌진흥청 축산연구소 가축유전자원시험장) ;
  • 최선호 (농촌진흥청 축산연구소 가축유전자원시험장) ;
  • 조창연 (농촌진흥청 축산연구소 가축유전자원시험장) ;
  • 한만희 (농촌진흥청 축산연구소 가축유전자원시험장) ;
  • 손동수 (농촌진흥청 축산연구소 가축유전자원시험장) ;
  • 이규승 (충남대학교 동물자원학부)
  • Published : 2004.12.01
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Abstract

This study was carried out to examine the effects of development media and those change on in vitro development (IVD) of porcine oocytes matured and fertilized in vitro. Putative embryos, which were matured in vitro in modified NCSU-23 (mNCSU-23) supplemented with 10％ porcine follicular fluid (pFF) and were fertilized in mTBM, were developed in vitro as the experimental scheme. The results are as follows. When porcine putative embryos were cultured in vitro in NCSU-23 or CZB/Pig-MEM, the percentage of oocytes cleaved was not different between two systems, but the percentage of blastocysts in NCSU-23 was significantly higher than in CZB/Pig-MEM (P<0.05). And when porcine putative embryos were cultured in vitro in NCSU-23 during 7 days with or without changing media at day 5, which was supplement with or without 10％ fetal bovine serum(FBS), the percentage of oocytes cleaved, blastocysts at day 6, and the cell number of ICM, TE and total of blastocysts at day 7 were not different among three treatments. As a result of this study, it is supposed that NCSU-23 be more favorable, to develop porcine embryos derived from IVM/IVF, than CZB/Pig-MEM, but that demonstration on the effects of changing medium with fresh one stand in need of the more experiments.

본 연구는 체외성숙/체외수정 유래의 돼지 난자를 이용하여 체외발달시 배양액의 종류나 교체에 따른 영향을 구명하고자 수행하였다. mNCSU-23에서 체외성숙시킨 다음 mTBM에서 체외수정시킨 난자를 목적에 따라 두 가지로 나누어 실험한 결과는 다음과 같다. 1. 체외성숙/체외수정란을 NCSU-23에서 배양액 교체없이 7일 동안 배양하거나 CZB에서 4일 배양한 다음 Pig-MEM으로 옮겨서 나머지 3일간 배양한 결과, 난분할율은 배양액간 차이를 보이지 않은 반면, 추정수정란대 배반포 발달을(P<0.05)과 분할란대 배반포 발달율은 NCSU-23에서 배양된 것이 유의적으로 높았다(P<0.01). 그러나 배 반포의 ICM 세포수, TE 세포수 및 총세포수에서는 모두 차이가 없었다. 2. 체외성숙/체외수정란를 NCSU-23에서 배양액 교체없이 7일 동안 배양하거나 체외배양 5일째에 신선한 동일 배양액이나 0.4％ BSA를 10％ FBS로 대체한 배양액(mNCSU-23F)으로 완전히 교체하여 배양한 결과, 난분할율, 배반포발달율, 배반포의 ICM 세포수, TE 세포수 및 총세포수 모두 처리간 유의적인 차이를 보이지 않았다. 결과적으로, NCSU-23이 CZB/Pig-MEM보다 체외성숙/체외수정 유래의 난자를 체외발달시키는데 적합한 것으로 사료되며, 체외발달배양 과정에 신선한 배양액이나 일부 변경된 배양액으로의 교체에 대해서는 더 많은 연구가 필요할 것으로 사료된다.

Keywords

References

  1. Abeydeera LR. 2001. In vitro fertilization and embryo development in the pig. J. Reprod. Fertil., 58(Suppl.):159-173
  2. Abeydeera LR. 2002. In vitro production of embryos in swine. Theriogenology, 57:256-273
  3. Barnett DK and Bavister BD. 1992. Hypotaurine requirement for in vitro development of golden hamster one-cell embryos into morulae and blastocysts, and production of term offspring from in vitro-fertilized ova. Biol. Reprod., 47:297-304 https://doi.org/10.1095/biolreprod47.2.297
  4. Bavister BD and Arlotto T. 1990. Influence of single amino acids on the development of hamster one-cell embryos in vitro. Mol. Reprod. Dev., 25:45-52 https://doi.org/10.1002/mrd.1080250109
  5. Bormann CL, Ongeri EM and Krisher RL. 2000. Effects of vitamins on maturation of caprime oocytes and their subsequent developmental capacity in vitro. Biol. Reprod., 62(Suppl.):166 (abstr.)
  6. Chatot CL, Ziomek CA, Bavister BD, Lewis JL and Torres I. 1989. An improved culture medium supports development of random-bred 1- cell mouse embryos in vitro. J. Reprod. Fertil., 86:679-688 https://doi.org/10.1530/jrf.0.0860679
  7. Davis DL. 1985. Culture and storage of pig embryos. J. Reprod. Fertil., 33(Suppl.):115-124
  8. Day BN. 2000. Reproductive biotechnologies: current status in porcine reproduction. Anim. Reprod. Sci., 60: 161-172 https://doi.org/10.1016/S0378-4320(00)00079-8
  9. Day BN, Abeydeera LR, Johnson LA, Welch GR, Wang WH, Cantley TC and Rieke A. 1998. Birth of piglets preselected for gender following in vitro fertilization of in vitro matured pig oocytes by X and Y bearing spermatozoa stored by high speed flow cytometry. Theriogenology, 49:360(abstr.) https://doi.org/10.1016/S0093-691X(98)90713-0
  10. Dobrinsky JR, Johnson LA and Rath D. 1996. Development of a culture medium (BECM-3) for porcine embryos: effects of bovine serum albumin and fetal bovine serum on embryo development. Biol. Reprod., 55:1069-1074 https://doi.org/10.1095/biolreprod55.5.1069
  11. Gandhi AP, Lane M, Gardner DK and Krisher RL. 2001. Substrate utilization in porcine embryos cultured in NCSU23 and G1.2/G2.2 sequential culture media. Mol. Reprod. Dev., 58:269-275 https://doi.org/10.1002/1098-2795(200103)58:3<269::AID-MRD4>3.0.CO;2-L
  12. Gardner DK and Lane M. 1993. Amino acids and ammonium regulate mouse embryo development in culture. Biol. Reprod., 48:377-385 https://doi.org/10.1095/biolreprod48.2.377
  13. Gardner DK, Schoolcraft WB, Wagley L, Schlenker T, Stevens J and Hesla J. 1998. A prospective randomized trial of blastocyst culture and transfer in in-vitro fertilization. Human Reprod., 13:3434-3440 https://doi.org/10.1093/humrep/13.12.3434
  14. Iwasaki T, Kimura E and Totsukawa K. 1999. Studies on a chemically defined medium for in vitro culture of in vitro matured and fertilized porcine oocytes. Theriogenology, 51:709-720 https://doi.org/10.1016/S0093-691X(99)00020-5
  15. Koo DB, Kim NH, Lee HT and Chung KS. 1997. Effects of fetal calf serum, amino acids, vitamins and insulin on blastocoel formation and hatching of in vivo and IVM/IVF-derived porcine embryos developing in vitro. Theriogenology, 48:791-802 https://doi.org/10.1016/S0093-691X(97)00302-6
  16. Krisher RL, Lane M and Bavister BD. 1993. Developmental competence and metabolism of bovine embryos cultured in semi defined and defined culture media. Biol. Reprod., 60:1345-1352 https://doi.org/10.1095/biolreprod60.6.1345
  17. Liu Z. and Foote RH. 1995. Effects of amino acids on the development of in-vitro matured/in-vitro fertilization bovine embryos in a simple protein-free medium. Human Reprod., 10:2985- 2991 https://doi.org/10.1093/oxfordjournals.humrep.a135834
  18. Liu Z. and Foote RH. 1997. Effects of amino acids and amanitin on bovine embryo development in a simple protein-free medium. Mol. Reprod. Dev., 46:278-285 https://doi.org/10.1002/(SICI)1098-2795(199703)46:3<278::AID-MRD6>3.0.CO;2-M
  19. Long CR, Dobrinsky JR and Johnson LA. 1999. In vitro production of pig embryos: Comparisons of culture media and boars. Theriogenology, 51:1375- 1390 https://doi.org/10.1016/S0093-691X(99)00081-3
  20. 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
  21. Malayer JR, Hansen PJ and Buhi WC. 1988. Secretion of proteins by cultured bovine oviducts collected from estrus through early diestrus. J. Exp. Zool., 248:345-353 https://doi.org/10.1002/jez.1402480313
  22. Marchal R, Feugang JM, Perreau C, Venturi E, Terqui M and Mermillod P. 2001. Meiotic and developmental competence of prepubertal and adult swine oocytes. Theriogenology, 56:17-29 https://doi.org/10.1016/S0093-691X(01)00539-8
  23. Mattioli M, Bacci ML, Galeati G and Seren E. 1989. Developmental competence of pig oocytes matured and fertilized in vitro. Theriogenology, 46:1201-1207
  24. Motlik J and Fulka J. 1974. Fertilization of pig follicular oocytes cultivated in vitro. J. Reprod. Fertil., 36:235-237 https://doi.org/10.1530/jrf.0.0360235
  25. Petters RM and Reed ML. 1991. Addition of taurine or hypotaurine to culture medium improves development of one- and two-cell pig embryos in vitro. Theriogenology, 35:253(abstr.) https://doi.org/10.1016/0093-691X(91)90229-7
  26. Petters RM and Wells KD. 1993. Culture of pig embryos. J. Reprod. Fertil., 48:61-73
  27. Pollard JW, Plante C and Leibo SP. 1995. Comparison of development of pig zygotes and embryos in simple and complex culture media. J. Reprod. Fertil., 103:331-337 https://doi.org/10.1530/jrf.0.1030331
  28. Rath D, Niemann H, Tao T and Boerjan M. 1995. Ratio and number of inner cell mass and trophoblast cells of in vitro and in vivo produced porcine embryos. Theriogenology, 43:304(abstr.) https://doi.org/10.1016/0093-691X(95)92458-L
  29. Stewart-Savage J and Bavister BD. 1988. Dete-rioration of stored culture media as monitored by a sperm motility bioassay. J. In Vitro Fertil. Embryo Transf., 5:76-80 https://doi.org/10.1007/BF01130662
  30. Swain JE, Bormann CL and Krisher RL. 2001. Development and viability of in vitro derived porcine blastocysts cultured in NCSU23 and G1.2/G2.2 sequential medium. Theriogenology, 56: 459-469 https://doi.org/10.1016/S0093-691X(01)00577-5
  31. Yoshida M, Mizoguchi Y, Ishigaki K, Kojima T and Nagai T. 1993. Birth of piglets derived from in vitro fertilization of pig oocytes matured in vitro. Theriogenology, 39:1303-1311 https://doi.org/10.1016/0093-691X(93)90232-T
  32. Youngs CR, Ford SP, McGinnis LK and Anderson LH. 1993. Investigations into the control of litter size in swine: I. Comparative studies on in vitro development of Meishan and Yorkshire preimplantation embryos. J. Anim. Sci., 71: 1561-1565 https://doi.org/10.2527/1993.7161561x
  33. 연성흠, 손동수, 한만희, 위미순, 최선호, 이규승. 2004a. 돼지 난포란 유래 체외수정란 생산에 대한 제요인의 영향. Ⅱ. 체외성숙배양시 EGF와 COC의 수가 체외성숙, 체외수정 및 체외발달에 미치는 영향. 한국수정란이식학회지, 19: 173-183
  34. 연성흠, 최선호, 김종대, 손동수, 한만희, 이규승. 2004b. 돼지 난포란 유래 체외수정란 생산에 대한 제요인의 영향. Ⅰ. 체외성숙, 체외수정, 체외발달에 대한 체외성숙 배양액의 영향. 한국수정란이식학회지, 19:165-172