Reproductive and Developmental Biology

The Korean Society of Animal Reproduction (한국동물번식학회)
권호 표지

Production of Cloned Jeju Black Cattle (Korean Cattle) from SCNT Embryo using Vitrification, One-Step Dilution and Direct Transfer Technique

초자화 동결과 1-단계 융해된 체세포 핵이식란의 직접 이식 기술로 제주흑우 복제소 생산

  • 김은영 (미래생명공학연구소) ;
  • 박민지 (미래생명공학연구소) ;
  • 김재연 (미래생명공학연구소) ;
  • 박효영 (미래생명공학연구소) ;
  • 노은지 (미래생명공학연구소) ;
  • 노은형 (미래생명공학연구소) ;
  • 송동환 ;
  • 오창언 (제주대학교 줄기세포연구센터) ;
  • 김영훈 (제주특별자치도 축산진흥원) ;
  • 문성호 (제주대학교 줄기세포연구센터) ;
  • 이동선 (제주대학교 생명자원과학대학 생명공학부) ;
  • 고문석 (국립축산과학원 난지축산시험장) ;
  • 류기중 (제주대학교 생명자원과학대학 생명공학부) ;
  • 박세필 (미래생명공학연구소)
  • Received : 2011.02.25
  • Accepted : 2011.03.03
  • Published : 2011.03.31
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Abstract

One-step dilution and direct transfer would be a practical technique for the field application of frozen embryo. This study was to examine whether Jeju Black Cattle (JBC, Korean Cattle) can be successfully cloned from vitrified and one-tep diluted somatic cell nuclear transfer (SCNT) blastocyst after direct transfer. For vitrification, JBC-SCNT blastocysts were serially exposed in glycerol (G) and ethylene glycol (EG) mixtures [10%, (v/v) G for 5 min., 10% G plus 20% EG (v/v) for 5 min., and 25% G plus 25% EG (v/v) for 30 sec.] which is diluted in 10% FBS added D-PBS. And then SCNT blastocysts were loaded in 0.25 ml mini straw, placed in cold nitrogen vapor for 3 min. and then plunged into $LN_2$. One-step dilution in straw was done in $25^{\circ}C$ water for 1 min, by placing vertically in the state of plugged-end up and down for 0.5 min, respectively. When in vitro developmental capacity of vitrified SCNT blastocyst was examined at 48 h after one-step dilution, hatched rate (56.4%) was slightly lower than that of control group (62.5%). In field trial, when the vitrified-thawed SCNT blastocysts were transferred into uterus of synchronized 5 recipients, a cloned female JBC was delivered by natural birth on day 299 and healthy at present. In addition, when the short tandem repeat marker analysis of the cloned JBC was evaluated, microsatellite loci of 11 numbers was perfectly matched genotype with donor cell (BK94-14). This study suggested that our developed vitrification and one-step dilution technique can be applied effectively on field trial for cloned animal production, which is even no longer in existence.

Keywords

References

  1. Baguisi A, Behboodi E, Melican DT, Pollock JS, Destrempes MM, Cammuso C, Williams JL, Nims SD, Porter CA, Midura P et al. (1999): Production of goats by somatic cell nuclear transfer. Nat Biotech 17:456-461. https://doi.org/10.1038/8632
  2. Chesne P, Adenot PG, Viglietta C, Baratte M, Boulanger L, Renard JP (2002): Cloned rabbits produced by nuclear transfer from adult somatic cells. Nat Biotech 20:336-369.
  3. Cibelli JB, Stice SL, Goleuke PJ, Kane JK, Jerry J, Balackwell C, de Leon A, Robl JM (1998): Cloned transgenic calves produced from nonquiescent fetal fibroblasts. Science 280:1256-1258. https://doi.org/10.1126/science.280.5367.1256
  4. Dochi O, Yamamoto Y, Saga H, Yoshiba N, Kano N, Maeda J, Miyata K, Yamauchi A, Tominaga K, Oda Y, Nakashima T, Inohae S (1998): Direct transfer of bovine embryos frozen-thawed in the presence of prophylene glycol or ethylene glycol under on-farm conditions in an integrated embryo transfer program. Theriogenology 49:1051-1058. https://doi.org/10.1016/S0093-691X(98)00053-3
  5. Gong G, Dai Y, Fan B, Zhu H, Zhu S, Wang H, Wang L, Tang B, Li R, Wan R, Liu Y, Huang Y, Zhang L, Sun X, Li N (2004): Birth of calves expressing the enhanced green fluorescent protein after transfer of fresh or vitrified/thawed blastocysts produced by somatic cell nuclear transfer. Mol Reprod Dev 69:278-288. https://doi.org/10.1002/mrd.20130
  6. Heyman Y, Chavatte-Palmer P, LeBourhis D, Camous S, Vignon X, Renard JP (2002): Frequency and occurrence of late-gestation losses from cattle cloned embryos. BioI Reprod 66:6-13. https://doi.org/10.1095/biolreprod66.1.6
  7. Im GS, Yang BS, Park SJ, Im SK, Yang BC, Yi YJ, Park CS (2001): Effect of protein supplimentation, $O_2$ concentration and co culture on the development of embryos produced by nuclear transfer using cultured cumulus cells in Hanwoo (Korean cattle). Asian-Aust J Anim Sci 14(9):1260-1266.
  8. Kato Y, Tani T, Sotomaru Y, Kurokawa K, Kato J, Doguchi H, Yasue H, Tsunoda Y (1998): Eight calves cloned from somatic cells of a single adult. Science 282:2095-2098. https://doi.org/10.1126/science.282.5396.2095
  9. Kim EY, Kim DI, Park NH, Weon YS, Nam HK, Lee KS, Park SY, Yoon SH, Park SP, Chung KS, Lim JH (1999): Systems for production of calves from Hanwoo (Korean Cattle) IVM/IVF/IVC blastocyst III. Vitrification and one-step dilution of Hanwoo blastocyst. Korean J Animal Reprod. 23(4):293-301.
  10. Kim EY, Kim YO, Kim JY, Park MJ, Park HY, Han YJ, Mun SH, Oh CE, Kim YH, Lee SS, Ko MS, Park SP (2010): In vitro development of somatic cell nuclear transfer embryo treated with flavonoid and production of cloned Jeju Black Cattle. Reprod Dev BioI 34(3):127-134.
  11. Lonergan ACO, Evans E, Boland D, Rozos T, Fair P, Duffy L-Y, Sung F, Du S, Chaubal J, Xu X, Yang, Tian XC (2007): Pregnancy and fetal characteristics after transfer of vitrified in vivo and cloned bovine embryos. Theriogenology 68:1128-1137. https://doi.org/10.1016/j.theriogenology.2007.08.012
  12. Massip A, Van der Zwalmen P, Leroy F (1984) Effect of stage of development on survival of mouse embryos frozen-thawed rapidly. Cryobiology 21:574-577. https://doi.org/10.1016/0011-2240(84)90057-9
  13. Park SP (2008) Industrialization and mass production of Jeju Black Cattle. Annual report on Korea Institute of Planning & Evaluation for Tedmology in Food, Agriculture, Forestry & Fisheries.
  14. Polejaeva lA, Chen SH, Vaught TD, Page RL, Mullins J, Ball S, Dai Y, Boone J, Walker S, Ayares DL et al. (2000): Cloned pigs produced by nuclear transfer from adult somatic cells. Nature 407:86-90. https://doi.org/10.1038/35024082
  15. RaIl WF, 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
  16. Shin T, Kraemer D, Pryor J, Liu L, Rugila J, Howe L, Buck S, Murphy K, Lyons L, Westhusin M (2002): A cat cloned by nuclear transplantation. Nature 415: 859. https://doi.org/10.1038/nature723
  17. Suzuki T, Yamamoto M, Ooe M, Sakata A, Matsuoka K, Nishikata Y, Okamoto K (1990): Effect of sucrose concentration used for one-step dilution upon in vitro and in vivo survival of bovine embryos refrigerated in glycerol and 1,2-propanediol. Theriogenology 34:1051-1057. https://doi.org/10.1016/S0093-691X(05)80004-4
  18. Tecirlioglu RT, French AJ, Lewis IM, Vajta G, Koorfiatis NA, Hall VJ, Ruddock NT, Cooney MA, Trounson O (2003): Birth of a cloned calf derived from a vitrified hand-made cloned embryo. Reprod Fert Dev 15:361-366. https://doi.org/10.1071/RD03052
  19. Voelkel SA, Hu YX (1992): Use of ethylene glycol as a cryoprotectant for bovine embryos allowing direct transfer of frozen-thawed embryos to recipient female. Theriogenology 37:687-697. https://doi.org/10.1016/0093-691X(92)90148-K
  20. Wakayama T, Perry AC, Zuccotti M, Johnson KR, Yanagimachi R (1998): Full-term development of mice from enucleated oocytes injected with cumulus cell nuelei. Nature 394:369-374. https://doi.org/10.1038/28615
  21. Wilmut I, Schnieke AE, McWhir J, Kind AJ, Campbell KH (1997): Viable offspring derived from fetal and adult mammalian cells. Nature 385:810-813. https://doi.org/10.1038/385810a0
  22. Yang BC, Im GS, Kim DH, Ko YG, Hwang SS, Nho WG, Kim MJ, Yang BS, Lee SJ, Seong HH (2008): Effects of gestation length and birth weight on survival rate in cloned Korean Native Calves. Reprod Dev BioI 32(1): 51-58.