DOI QR코드

DOI QR Code

Development of Bovine Nuclear Transfer Embryos Using Life-span Extended Donor Cells Transfected with Foreign Gene

  • Hwang, Seongsoo (Institute of Genetic Engineering, Hankyong National University) ;
  • Choi, Eun Joo (Institute of Genetic Engineering, Hankyong National University) ;
  • You, Seungkwon (College of Life and Environmental Sciences, Korea University) ;
  • Choi, Yun-Jaie (School of Agricultural Biotechnology, Seoul National University) ;
  • Min, Kwan-Sik (The Graduate School of Bio- and Information Technology) ;
  • Yoon, Jong-Taek (Institute of Genetic Engineering, Hankyong National University)
  • Received : 2005.12.26
  • Accepted : 2006.04.05
  • Published : 2006.11.01

Abstract

This study was performed to determine the developmental potentials of nuclear transfer (NT) embryos using life-span extended cells transfected with a foreign gene as donor cells. A life-span extended bovine embryonic fibroblast cell line was transfected with an expression vector in which the human type II collagen (BOMAR) and ear fibroblasts were used as a donor cell. Cytogenetic analysis was performed to analyze the chromosomal abnormality of donor cells. The fusion rate of 1.8 kV/cm for $15{\mu}sec$ given twice was significantly higher than that of other groups (p<0.05) and the embryos lysed were significantly higher after 1.8 kV/cm for $20{\mu}sec$ given once compared to other groups (p<0.01). The blastocyst development in the ear cell group was statistically significant compared to both BOMAR groups (p<0.01). Both BOMAR groups cultured more than 40 passages (>40 passages) had a lower number of chromosomes; however, fresh granulosa cell (GC) and BOMAR groups cultured less than 20 passages had normal chromosome numbers. Both >40 passages BOMAR groups had numerous obscure debris in metaphase spreads. The transfected foreign gene was expressed in all BOMAR groups, but not in the GC group. Based on these results, the lower developmental potential of NT embryos using life-span extended donor cells transfected with a foreign gene might be a cause of chromosomal abnormality in donor cells.

Keywords

Chromosomal Abnormality;Foreign Gene Transfection;Life-span Extended Donor Cell;Somatic Cell Nuclear Transfer

Acknowledgement

Supported by : Rural Development Administration

References

  1. Campbell, K. H. S., J. McWhir, W. A. Ritchie and I. Wilmut. 1996. Sheep cloned by nuclear transfer from a cultured cell line. Nature 380:64-66 https://doi.org/10.1038/380064a0
  2. Cibelli, J. B., S. L. Stice, P. J. Goleuke, J. J. Kane, J. Jerry, C. Blackwell, F. A. Ponce de Leon and J. M. Robl. 1998. Cloned transgenic calves produced from nonquiscent fetal fibroblasts. Sci. 280:1256-1258 https://doi.org/10.1126/science.280.5367.1256
  3. Hill, J. R., Q. A. Winger, C. R. Long, C. R. Looney, J. A. Thompson and M. E. Westhusin. 2000. Development rates of male bovine nuclear transfer embryos derived from adult and fetal cells. Biol. Reprod. 62:1135-1140 https://doi.org/10.1095/biolreprod62.5.1135
  4. Wakayama, T., A. C. Perry, M. Zuccotti, K. R. Johnson and R. Yanagimachi. 1998. Full-term development of mice from enucleated oocytes injected with cumulus cell nuclei. Nature 394:369-374 https://doi.org/10.1038/28615
  5. Zakhartchenko, V., R. Alberio, M. Stojkovic, K. Prelle, W. Schernthaner, P. Stojkovic, H. Wenigerkind, R. Wanke, M. Duchler, R. Steinborn, M. Mueller, G. Brem and E. Wolf. 1999. Adult cloning in cattle: Potential of nuclei from a permanent cell line and from primary cultures. Mol. Reprod. Dev. 54:264-272 https://doi.org/10.1002/(SICI)1098-2795(199911)54:3<264::AID-MRD7>3.0.CO;2-Y
  6. Kubota, C., H. Yamakuchi, J. Todoroki, K. Mizoshita, N. Tabara, M. Barber and X. Yang. 2000. Six cloned calves produced from adult fibroblast cells after long-term culture. Proc. Natl. Acad. Sci. USA. 97:990-995
  7. King, W. A. 1990. Chromosome abnormalities and pregnancy failure in domestic animals. Adv. Vet. Sci. Comp. Med. 34:229-250
  8. Li, G. P., K. L. White, K. I. Aston, L. N. Meerdo and T. D. Bunch. 2004. Conditioned medium increases the polyploid cell composition of bovine somatic cell nuclear-transferred blastocysts. Reprod. 127:221-228 https://doi.org/10.1530/rep.1.00089
  9. Dong, Y. J., R. Rajamahendran, T. Otoi, N. R. Mtango, M. D. Varisanga, X. J. Bai and T. Suzuki. 2004. Production of cloned calves by the transfer of somatic cells derived from frozen tissues using simple portable $CO_2$ incubator. Asian-Aust. J. Anim. Sci. 17:168-173 https://doi.org/10.5713/ajas.2004.168
  10. Arat, S., J. Gibbons, A. J. Rzucidleo, D. S. Respess, M. Tumlin and S. L. Stice. 2002. In vitro development of bovine nuclear transfer embryos from transgenic clonal lines of adult and fetal fibroblast cells of the same genotype. Biol. Reprod. 66:1768-1774 https://doi.org/10.1095/biolreprod66.6.1768
  11. Hanada, H., K. Takeda, T. Tagami, K. Nirasawa, S. Akagi, N. Adachi, S. Takahashi, Y. Izakei, M. Iwamoto, D. Fuchimoto, N. Miyashita, M. Kubo, A. Onishi and W. A. King. 2005. Chromosomal instability in the cattle clones derived by somatic cell nuclear-transfer. Mol. Reprod. Dev. 71:36-44 https://doi.org/10.1002/mrd.20283
  12. Collas, P., C. Pinto-Correia, F. A. Ponce de Leon and J. M. Robl. 1992. Effect of donor cell cycle stage on chromatin and spindle morphology in nuclear transplant rabbit embryos. Biol. Reprod. 46:501-511 https://doi.org/10.1095/biolreprod46.3.501
  13. Polejaeva, I. A., S. H. Chen, T. D. Vaught, R. L. Page, J. Mullins, S. Ball, Y. Dai, J. Boone, S. Walker, D. L. Ayares, A. Colman and K. H. S. Campbell. 2000. Cloned pigs produced by nuclear transfer from adult somatic cells. Nature 407:86-90 https://doi.org/10.1038/35024082
  14. Schnieke, A. E., A. J. Kind, W. A. Ritchie, T. Mycock, A. R. Scott, M. Ritchie, I. Wilmut, A. Colman and K. H. S. Campbell. 1997. Human factor IX transgenic sheep produced by transfer of nuclei from transfected fetal fibroblasts. Sci. 278:2130-2133 https://doi.org/10.1126/science.278.5346.2130
  15. Slimane-Bureau, W., V. Bordignon, C. Leveillee, L. C. Smith and W. A. King. 2003. Assessment of chromosomal abnormalities in bovine nuclear transfer embryos and in their donor cells. Cloning Stem Cells 5:123-133 https://doi.org/10.1089/153623003322234722
  16. Booth, P. J., D. Viuff, S. Tan, P. Holm, T. Greve and H. Callesen. 2003. Numerical chromosome errors in day 7 somatic nuclear transfer bovine blastocysts. Biol. Reprod. 68:922-928 https://doi.org/10.1095/biolreprod.102.007187
  17. Campbell, K. H. S., W. A. Ritchie and I. Wilmut. 1993. Nuclearcytoplasmic interactions during the first cell cycle of nuclear transfer reconstructed bovine embryos: Implications for deoxyribonucleic acid replication and development. Biol. Reprod. 49:933-942 https://doi.org/10.1095/biolreprod49.5.933
  18. Chen, S. H., T. D. Vaught, J. A. Monahan, J. Boone, E. Emslie, P. M. Jobst, A. E. Lamborn, A. Schnieke, L. Robertson, A. Colman, Y. Dai, I. A. Polejaeva and D. L. Ayares. 2002. Efficient production of transgenic cloned calves using preimplantation screening. Biol. Reprod. 67:1488-1492 https://doi.org/10.1095/biolreprod.102.006981
  19. Onishi, A., M. Iwamoto, T. Akita, S. Mikawa, K. Takeda, T. Awata, H. Hanada and A. C. F. Perry. 2000. Pig cloning by microinjection of fetal fibroblast nuclei. Sci. 289:1188-1190 https://doi.org/10.1126/science.289.5482.1188
  20. Bureau, W. S., V. Bordignon, C. Leveillee, L. C. Smith and W. A. King. 2003. Assessment of chromosomal abnormalities in bovine nuclear transfer embryos and in the donor cells. Cloning Stem Cells 5:123-132 https://doi.org/10.1089/153623003322234722
  21. Cui, W., D. Wylie, S. Aslam, A. Dinnyes, T. King, I. Wilmut and A. J. Clark. 2003. Telomerase-immortalized sheep fibroblasts can be reprogrammed by nuclear transfer to undergo early development. Biol. Reprod. 69:15-21 https://doi.org/10.1095/biolreprod.102.013250
  22. Zakhartchenko, V., S. Mueller, R. Alberio, W. Schernthaner, M. Stojkovic, H. Wenigerkind, R. Wanke, C. Lassnig, M. Mueller, E. Wolf and G.. Brem. 2001. Nuclear transfer in cattle with non-transfected and transfected fetal or cloned transgenic fetal and postnatal fibroblasts. Mol. Reprod. Dev. 60:362-329 https://doi.org/10.1002/mrd.1098
  23. Mohamed, N. M. S. and Y. Takahashi. 2000. In vitro developmental potential of bovine nuclear transfer embryos derived from primary cultured cumulus cells. J. Vet. Med. Sci. 62:339-342 https://doi.org/10.1292/jvms.62.339
  24. You, S. K., M. H. Heo, J. H. Moon, S. C. Kim, S. W. Kwak, D. H. Yoon, D. I. Jin, K. C. Hong, D. N. Foster, Y. J. Choi and H. G. Lim. 2004. Establishment of life-span extended bovine fibroblast cells carrying the characterization of primary cells. Mol. Cells. 18:261-268
  25. Shin, T., D. Kraemer, J. Pryor, L. Liu, J. Rugila, L. Howe, S. Buck, K. Murphy, L. Lyons and M. Westhusin. 2002. A cat cloned by nuclear transplantation. Nature 415:859 https://doi.org/10.1038/nature723
  26. Lee, B. C., M. K. Kim, G. Jang, H. J. Oh, F. Yuda, H. J. Kim, M. H. Shamim, J. J. Kim, S. K. Kang, G. Schatten and W. S. Hwang. 2005. Dogs cloned from adult somatic cells. Nature 436:641 https://doi.org/10.1038/436641a
  27. Kato, Y., T. Tani, Y. Sotomaru, K. Kurokawa, J. Kato, H. Doguchi, H. Yasue and Y. Tsunoda. 1998. Eight calves cloned from somatic cells of a single adult. Sci. 282:2095-2098 https://doi.org/10.1126/science.282.5396.2095
  28. Galli, C., I. Lagutina, G. Crotti, S. Colleoni, P. Turini, N. Ponderato, R. Duchi and G. Lazzari. 2003. A cloned horse born to its dam twin. Nature 424:635
  29. Hong, S. B., S. J. Uhm, H. Y. Lee, C. Y. Park, M. K. Gupta, B. H. Chung, K. S. Chung and H. T. Lee. 2005. Developmental ability of bovine embryos nuclear transferred with frozenthawed or cooled donor cells. Asian-Aust. J. Anim. Sci. 18:1242-1248 https://doi.org/10.5713/ajas.2005.1242