Skip to main content
SpringerLink
Log in

Production of transgenic blastocyst of sheep by somatic cell cloning

  • Notes
  • Published:
Chinese Science Bulletin

Abstract

Five samples from primary cultures of five sheep ovarian granulosa cells were transfected by pEGFP-N1 DNA. Five transgenic positive cell lines, each from one of the five samples above, were used as donor nuclei for somatic nucleus transfer. A total of 352in vitro matured and enucleated sheep oocytes were fused electrically with transgenic granulosa cells and 329 reconstructed embryos were obtained after activation by Ionomycin/6-DMAP, and these embryos were cultured in SOFaaBSA medium for 7 d. The result shows that 312 embryos (94.8%) had gone through cleavage and among them 63 (19.1%) had developed to the blastocyst stage. Expression of GFP gene was detected in various stages of early embryonic development by sampling randomly. Blastocyst rates given by the four cells treated with 0.5% FCS starvation was 19.6% (55/280) and it had not shown difference significantly (P>0.05) with the result obtained with another cell line that had not gone through serum starvation (16.3%, 8/49). This experiment indicates that sheep transgenic embryos up to the blastocyst stage can be produced effectively by the combination of gene transfection in somatic cells in culture and somatic cell cloning.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Wilmut, I., Schnieke, A. E., McWhir, J. et al., Viable offspring derived from fetal and adult mammalian cells, Nature, 1997, 385: 810.

    Article  Google Scholar 

  2. Wakayama, T., Perry, A. C. F., Zuccotti, M. et al., Full-term development of mice from enucleated oocytes injected with cumulus cell nuclei, Nature, 1998, 394: 369.

    Article  Google Scholar 

  3. Cibelli, J. B., Stice, S. T., Golueke, P. J. et al., Cloned transgenic calves produced from nonquiescent fetal fibroblasts, Science, 1998, 280: 1256.

    Article  Google Scholar 

  4. Kato, Y., Tani, T., Sotomaru, Y. et al., Eight calves cloned from somatic cells of a single adult, Science, 1998, 282: 2095.

    Article  Google Scholar 

  5. Baguisi, A., Behboodi, E., Melican, D. et al., Production of goats by somatic cell nuclear transfer, Nature Biotech., 1999, 17: 456.

    Article  Google Scholar 

  6. Polejaeva, I. A., Chen, S. H., Vaught, T. D. et al., Cloned pigs produced by nuclear transfer from adult somatic cells, Nature, 2000, 407: 86.

    Article  Google Scholar 

  7. Onishi, A., Iwamoto, M., Akita, T. et al., Pig cloning by microinjection of fetal fibroblast nuclei, Science, 2000, 289: 1188.

    Article  Google Scholar 

  8. Wakayama, T., Shinkai, Y., Tamashiro, K. L. K. et al., Cloning of mice to six generations, Nature, 2000, 407: 318.

    Article  Google Scholar 

  9. Cindy, T. X., Xu, J., Yang, X. Z., Normal telomere lengths found in cloned cattle, Nature Genet., 2000, 26: 272.

    Article  Google Scholar 

  10. Eggan, K., Akutsu, H., Hocheclinger, K. et al., X-chromosome inactivation in cloned mouse embryos, Science, 2000, 290: 1578.

    Article  Google Scholar 

  11. Schnieke, A. E., Kind, A. J., Ritchie, W. A. et al., Human factor IX transgenic sheep produced by transfer of nuclei from transfected fetal fibroblasts, Science, 1997, 278: 2130.

    Article  Google Scholar 

  12. McCreath, K. J., Howcroft, J., Campbell, K. H. S. et al., Production of gene-targeted sheep by nuclear transfer from cultured somatic cells, Nature, 2000, 405: 1066.

    Article  Google Scholar 

  13. Wells, D. N., Misica, P. M., Day, A. M. et al., Production of cloned lambs from an established embryonic cell line: A comparison betweenin vivo- andin vitro-matured cytoplasts, Biol. Reprod., 1997, 57: 385.

    Article  Google Scholar 

  14. Campbell, K. H. S., McWhir, J., Ritchie, W. A. et al., Sheep cloned by nuclear transfer from a cultured cell line, Nature, 1996, 380: 64.

    Article  Google Scholar 

  15. Tani, T., Kato, Y., Tsunoda, Y., Development potential of cumulus cell-derived cultured cells frozen in a quiescent state after nucleus transfer, Theriogenology, 2000, 53: 1623.

    Article  Google Scholar 

  16. Wassarman, P. M., Albertini, D. F., The mammalian ovum, in The Physiology of Reproduction (eds. Knobil, E., Neill, J. D.), New York: Raven Press, 1994, 79.

    Google Scholar 

  17. Prather, R. S., First, N. L., Cloning embryos by nuclear transfer, J. Reprod Fertil Suppl., 1990, 41: 125.

    Google Scholar 

  18. Campbell, K. H. S., Ritchie, W. A., Wilmut, I., Nuclear-cytoplasmic interactions during the first cell cycle of nuclear transfer reconstructed bovine embryos: implications for deoxyribonucleic acid replication and development, Biol. Reprod., 1993, 49: 933.

    Article  Google Scholar 

  19. Chastant, S., Christians, E., Campion, E. et al., Quantitative control of gene expression by nucleocytoplasmic interactions in early mouse embryos: consequences for reprogrammation by nuclear transfer, Mol. Reprod. Dev., 1996, 44: 423.

    Article  Google Scholar 

  20. Shiga, K., Fujita, T., Hirose, K. et al., Production of calves by transfer of nuclei from cultured somatic cells obtained from Japanese black bulls, Theriogenology, 1999, 52: 527.

    Article  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. State Key Laboratory for Agrobiotechnologies, China Agricultural University, 100094, Beijing, China

    Xiaorong An, Kemian Gou & Yongfu Chen

Authors
  1. Xiaorong An
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Kemian Gou
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Yongfu Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Kemian Gou.

Additional information

These authors contributed equally to this work.

About this article

Cite this article

An, X., Gou, K. & Chen, Y. Production of transgenic blastocyst of sheep by somatic cell cloning. Chin.Sci.Bull. 46, 1630–1634 (2001). https://doi.org/10.1007/BF02900623

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02900623

Keywords

Search

Navigation