Reproduction, Fertility and Development Reproduction, Fertility and Development Society
Vertebrate reproductive science and technology
RESEARCH ARTICLE

No differences in sheep somatic cell nuclear transfer outcomes using serum-starved or actively growing donor granulosa cells

T. T. Peura A C D , K. M. Hartwich A , H. M. Hamilton A B and S. K. Walker A
+ Author Affiliations
- Author Affiliations

A South Australian Research and Development Institute, Turretfield Research Centre, Rosedale, SA 5350, Australia.

B Department of Animal Science, The University of Adelaide Roseworthy Campus, Roseworthy, SA 5371, Australia.

C Present address: Sydney IVF Ltd, GPO Box 4384, Sydney, NSW 2001, Australia.

D To whom correspondence should be addressed. email: teija.peura@sivf.com.au

Reproduction, Fertility and Development 15(3) 157-165 https://doi.org/10.1071/RD02092
Submitted: 3 October 2002  Accepted: 6 May 2003   Published: 6 May 2003

Abstract

The aim of this study was to compare serum-starved and non-starved donor cells in sheep nuclear transfer with a special emphasis on cloning outcomes. Sheep oocytes, derived either in vivo or in vitro, were fused with cultured serum-starved or actively growing adult granulosa cells. Resulting blastocysts were transferred to recipients fresh or after vitrification, and subsequent pregnancies followed to term. Donor cell treatment did not significantly affect preimplantation development, pregnancy rates, fetal loss or neonate survival rates. Of 22 lambs born, ten survived the immediate perinatal period but all succumbed at various timepoints within the first few weeks of life. The results of the study suggest that the use of serum-starved cells offers no advantages or disadvantages to cloning outcomes. Neither were significant differences in outcomes observed when using either in vivo- or in vitro-derived oocytes or embryos transferred fresh or after vitrification. Yet, these results continue to highlight problems associated with somatic cell cloning as indicated by offspring mortality. It remains unclear whether the high offspring mortality in the current study was related to species, associated with the cell lines used or the result of other causes.

Extra keywords: cloning


Acknowledgments

We thank Ms Jen Kelly, Ms Skye Rudiger and Dr David Kleemann for their expert technical assistance in oocyte collections and embryology and Dr Lyn Davies for performing Caesarean sections. We also thank Mr Clive MacLaughlan and Dr Simon Bawden for their contribution to the microsatellite analyses. This work was part of a collaborative research effort with The Grange/Multiplex Constructions Pty, whose financial support is gratefully acknowledged.


References

Akagi, S. , Takahashi, S. , Adachi, N. , Sugawara, T. , Tozuka, H. , Toya, T. , Nirasawa, K. , Yamamoto, E. , Shimizu, M. , and Izaike, Y. (2002). Production of calves by nuclear transfer using non-cultured and cultured cumulus cells Theriogenology 57,(Abstract.)


Baguisi, A. , Behboodi, E. , Melican, D. , Pollock, J. , and Destrempes, M. , et al. (1999). Production of goats by somatic cell nuclear transfer Nature Biotechnol. 17, 456–61.
CrossRef |

Bielas, J. , and Heddle, J. (2000). Proliferation is necessary for both repair and mutation in transgenic mouse cells Proc. Natl Acad. Sci. USA 97, 11 391–6.
CrossRef |

Boquest, A. , Day, B. , and Prather, R. (1999). Flow cytometric cell cycle analysis of cultured porcine fetal fibroblast cells Biol. Reprod. 60, 1013–19.
PubMed |

Campbell, K. , Loi, P. , Cappai, P. , and Wilmut, I. (1994). Improved development to blastocyst of ovine nuclear transfer embryos reconstructed during the presumptive S-phase of enucleated activated oocytes Biol. Reprod. 50, 1385–93.
PubMed |

Campbell, K. , McWhir, J. , Ritchie, W. , and Wilmut, I. (1996). Sheep cloned by nuclear transfer from a cultured cell line Nature 380, 64–6.
CrossRef | PubMed |

Cho, J. , Lee, B. , Park, J. , Lim, J. , Shin, S. , Kim, K. , Lee, B. , and Hwang, W. (2002). Development of bovine oocytes reconstructed with different donor somatic cells with or without serum starvation Theriogenology 57, 1819–28.
CrossRef | PubMed |

Cibelli, J. , Stice, S. , Golueke, P. , Kane, J. , Jerry, J. , Blackwell, C. , Ponce de Leon, F. , and Robl, J. (1998). Cloned transgenic calves produced from non-quiescent fetal fibroblasts Science 280, 1256–8.
CrossRef | PubMed |

De Sousa, P. , King, T. , Harkness, L. , Young, L. , Walker, S. , and Wilmut, I. (2001). Evaluation of gestational deficiencies in cloned sheep fetuses and placentae Biol. Reprod. 65, 23–30.
PubMed |

Denning, C. S. B. , Ainslie, A. , Bracken, J. , Dinnyes, A. , and Fletcher, J. , et al. (2001). Deletion of the alpha(1,3)galactosyl transferase (GGTA1) gene and the prion protein (PrP) gene in sheep Nature Biotechnol. 19, 559–62.
CrossRef |

Edwards, L. , Peura, T. , Hartwich, K. , Rudiger, S. , McMillen, I. , and Walker, S. (2002). Circulating ACTH and cortisol concentrations are higher in cloned lambs in the first month of life Endocrinology 143, 3699–702.
CrossRef | PubMed |

Eggan, K. , Akutsu, H. , Loring, J. , Jackson-Grusby, L. , Klemm, M. , Rideout, W. , Yanagimachi, R. , and Jaenishc, R. (2001). Hybrid vigor, fetal overgrowth, and viability of mice derived by nuclear cloning and tetraplois embryo complementation Proc. Natl Acad. Sci. USA 98, 6209–14.
CrossRef | PubMed |

Hill, J. , Roussel, A. , Cibelli, J. , Edwards, J. , and Hooper, N. , et al. (1999a). Clinical and pathological features of cloned transgenic calves and fetuses (13 case studies) Theriogenology 51, 1451–65.
CrossRef | PubMed |

Hill, J. , Winger, Q. , Jones, K. , Keller, D. , King, W. , and Westhusin, M. (1999b). The effect of donor cell serum starvation and oocyte activation compounds on the development of somatic cloned embryos Cloning 1, 201–8.
CrossRef |

Hill, J. , Winger, Q. , Long, C. , Looney, C. , Thompson, J. , and Westhusin, M. (2000). Developmental rates of male bovine nuclear transfer embryos derived from adult and fetal cells Biol. Reprod. 62, 1135–40.
PubMed |

Humpherys, D. , Eggan, K. , Akutsu, H. , Friedman, A. , Hochedlinger, K. , Yanagimachi, R. , Lander, E. , Golub, T. , and Jaenisch, R. (2002). Abnormal gene expression in cloned mice derived from embryonic stem cell and cumulus cell nuclei Proc. Natl Acad. Sci. USA 99, 12 889–94.
CrossRef |

Kato, Y. , Tetsuya, T. , Somtomary, Y. , Kurokawa, K. , Kato, J. , Doguchi, H. , Yasue, H. , and Tsunoda, Y. (1998). Eight calves cloned from somatic cells of a single adult Science 282, 2095–8.
CrossRef | PubMed |

Keefer, C. , Keyston, R. , Lazaris, A. , Bhatia, B. , and Begin, I. (2002). Production of cloned goats after nuclear transfer using adult somatic cells Biol. Reprod. 66, 199–203.
PubMed |

Kubota, C. , Yamakuchi, H. , Todoroki, J. , Mizoshita, K. , Tabara, N. , Barber, M. , and Yang, X. (2000). Six cloned calves produced from adult fibroblast cells after long-term culture Proc. Natl Acad. Sci. USA 97, 990–5.
CrossRef | PubMed |

Kues, W. , Anger, M. , Carnwath, J. , Paul, D. , Motlik, J. , and Niemann, H. (2000). Cell cycle synchronization of porcine fetal fibroblasts: effects of serum deprivation and reversible cell cycle inhibitors Biol. Reprod. 62, 412–19.
PubMed |

Lanza, R. , Cibelli, J. , Blackwell, C. , Christofalo, V. , and Francis, M. (2000). Extension of cell life-span and telemere length in animals cloned from senescent somatic cells Science 288, 665–9.
CrossRef | PubMed |

Lanza, R. , Cibelli, J. , Faber, D. , Sweeney, R. , Henderson, B. , Nevala, W. , West, M. , and Wettstein, P. (2001). Cloned cattle can be healthy and normal Science 294, 1893–4.
CrossRef | PubMed |

Loi, P. , Clinton, M. , Barboni, B. , Fulka, J. , Cappai, P. , Feil, R. , Moor, R. , and Ptak, G. (2002). Nuclei of nonviable ovine somatic cells develop into lambs after nuclear transplantation Biol. Reprod. 67, 126–32.
PubMed |

McCreath, K. , Howcroft, J. , Campbell, K. , Colman, A. , Schnieke, A. , and Kind, A. J. (2000). Production of gene-targeted sheep by nuclear transfer from cultured somatic cells Nature 405, 1066–9.
CrossRef | PubMed |

Mitalipova, M. , Beyhan, Z. , Chang, C. , and First, N. (2002). Effect of different cell types and cell lines on development of bovine nuclear transfer embryos Theriogenology 57,(Abstract.)


National Health and Medical Research Council. (1997). Australian Code of Practice for the care and use of Animals for Scientific Purposes (Commonwealth Department of Health and Family services: Canberra.)

Oback, B. , and Wells, D. (2002a). Donor cells for nuclear cloning—many are called but few are chosen Cloning Stem Cells 4, 147–68.
CrossRef | PubMed |

Oback, B. , and Wells, D. (2002b). Practical aspects of donor cell selection for nuclear cloning Cloning Stem Cells 4, 169–174.
CrossRef | PubMed |

Ogonuki, N. , Inoue, K. , Yamamoto, Y. , Noguchi, Y. , and Tanemura, K. (2002). Early death of mice cloned from somatic cells Nat. Genet. 30, 253–4.
CrossRef | PubMed |

Palma, G. , Zakhartchenko, V. , Alberio, R. , Kaiser, G. , Aller, J. , and Alberio, R. (2002). Production of adult somatic cell clones from bovine granulosa cells cultured to full confluence Theriogenology 57,(Abstract.)


Peura, T. (2000). Serum starvation can cause excessive DNA damage in sheep fetal fibroblasts Theriogenology 55,(Abstract.)


Peura, T. , Hartwich, K. , Kelly, J. , Hamilton, H. , Kleemann, D. , Rudiger, S. , and Walker, S. (2002). Survival of fresh and vitrified sheep IVP and cloned embryos after embryo transfer. In ‘Proceedings of the Australian Society of Reproductive Biology, 22–25 September 2002, Adelaide, Australia.’ Reprod. Fertil. Dev. 14, 50.(Suppl.), (Abstract 16.)


Peura, T. , Kleemann, D. , Rudiger, S. , Nattrass, G. , McLaughlan, C. , and Walker, S. (2003). Effect of nutrition of oocyte donor on the outcomes of somatic cell nuclear transfer in the sheep Biol. Reprod. 68, 45–50.
PubMed |

Schnieke, A. , Kind, A. , Ritchie, W. , Mycock, K. , Scott, A. , Ritchie, M. , Wilmut, I. , Colman, A. , and Campbell, K. (1997). Human factor IX transgenic sheep produced by transfer of nuclei from transfected fetal fibroblasts Science 278, 2130–3.
CrossRef | PubMed |

Shin, T. , Kraemer, D. , Pryor, J. , Liu, L. , Rugila, J. , Howe, L. , Buck, S. , Murphy, K. , Lyons, L. , and Westhusin, M. (2002). A cat cloned by nuclear transplantation Nature 415,
CrossRef | PubMed |

Tani, T. , Kato, Y. , and Tsunoda, Y. (2001). Direct exposure of chromosomes to nonactivated ovum cytoplasm is effective for bovine somatic cell nucleus reprogramming Biol. Reprod. 64, 324–30.
PubMed |

Vajta, G. , Kuwayama, M. , Holm, P. , Booth, P. , Jacobsen, H. , Greve, T. , and Callesen, H. (1998). A new way to avoid cryoinjuries of mammalian ova and embryos: the OPS-vitrification Mol. Reprod. Dev. 51, 53–8.
CrossRef | PubMed |

Wakayama, T. , Perry, A. , Zuccotti, M. , Johnson, K. , and Yanagamachi, R. (1998). Full-term development of mice from enucleated oocytes injected with cumulus cell nuclei Nature 394, 369–74.
CrossRef | PubMed |

Walker, S. , Hill, J. , Kleemann, D. , and Nancarrow, C. (1996). Development of ovine embryos in synthetic oviductal fluid containing amino acids at oviductal fluid concentrations Biol. Reprod. 55, 703–8.
PubMed |

Wells, D. , Misica, P. , Day, T. , and Tervit, H. (1997). Production of cloned lambs from an established embryonic cell line; a comparison between in vivo and in vitro-matured cytoplasts Biol. Reprod. 57, 385–93.
PubMed |

Wells, D. , Misica, P. , and Tervit, H. (1999). Production of cloned calves following nuclear transfer with cultured adult mural granulosa cells Biol. Reprod. 60, 996–1005.
PubMed |

Wells, D. , Tucker, F. , Miller, A. , Oliver, J. , Xiang, T. , Forsyth, J. , Berg, M. , Cockrem, K. , and Tervit, H. (2002). Effect of early or late G1 cell cycle phase on embryo development following somatic cell nuclear transfer in cattle Theriogenology 57,(Abstract.)


Wilmut, I. , Schnieke, A. , McWhir, J. , Kind, A. , and Campbell, K. (1997). Viable offspring derived from fetal and adult mammalian cells Nature 385, 810–13.
CrossRef | PubMed |

Yin, X. , Tani, T. , Yonemura, I. , Kawakami, M. , Miyamoto, K. , Hasegawa, R. , Kato, Y. , and Tsunoda, Y. (2002). Production of cloned pigs from adult somatic cells by chemically assisted removal of maternal chromosomes Biol. Reprod. 67, 442–6.
PubMed |

Zakhartchenko, V. , Durcova-Hills, G. , Stojkovic, M. , Schernthaner, W. , Prelle, K. , Steinborn, R. , Müller, M. , Brem, G. , and Wolf, E. (1999). Effects of serum starvation and re-cloning on the efficiency of nuclear transfer using bovine fetal fibroblasts J. Reprod. Fertil. 115, 325–31.
PubMed |



Rent Article (via Deepdyve) Export Citation Cited By (14)