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.
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