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Effect of donor cell age on development of ovine nuclear transfer embryos in vitro

Published online by Cambridge University Press:  25 June 2010

B. Heidari ,
A. Shirazi ,
P. Tajic ,
E. Ahmadi ,
H. Nazari ,
N. Shams-Esfandabadi  and
H. Ghasemzadeh-Nava
B. Heidari
Affiliation:
Research Institute of Animal Embryo Technology, Shahrekord University, Shahrekord, Iran. Department of Veterinary Biomedical Sciences, Western College of Veterinary Medicine, University of Saskatchewan, Saskatoon, Canada.
A. Shirazi*
Affiliation:
Department of Veterinary Biomedical Sciences, Western College of Veterinary Medicine, University of Saskatchewan, 52 Campus Drive, Saskatoon SK S7N 5B4, Canada. Research Institute of Animal Embryo Technology, Shahrekord University, Shahrekord, Iran.
P. Tajic
Affiliation:
Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran.
E. Ahmadi
Affiliation:
Research Institute of Animal Embryo Technology, Shahrekord University, Shahrekord, Iran.
H. Nazari
Affiliation:
Research Institute of Animal Embryo Technology, Shahrekord University, Shahrekord, Iran.
N. Shams-Esfandabadi
Affiliation:
Research Institute of Animal Embryo Technology, Shahrekord University, Shahrekord, Iran.
H. Ghasemzadeh-Nava
Affiliation:
Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran.
*
All correspondence to: Abolfazl Shirazi. Department of Veterinary Biomedical Sciences, Western College of Veterinary Medicine, University of Saskatchewan, 52 Campus Drive, Saskatoon SK S7N 5B4, Canada. Tel: +1 306 966 4317. Fax: +1 306 966 7376. e-mail: shiraziabbas@ yahoo.com; a.shirazi@avicenna.ac.ir
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Summary

The effects of the age of cell donor animal on in vitro development of ovine nuclear transfer (NT) embryos were investigated. Somatic donor cells were obtained from two different sources: (1) adult cells (adult fibroblast cells; AFC and adult cumulus cells; ACC); and (2) fetal fibroblasts (40-day-old; FFC-40 and 65-day-old; FFC-65). The fibroblast cell lines were used for NT procedures within 4–13 subpassages. While the cumulus cells were used as non-cultured (fresh) cells. The in vitro matured abattoir-derived oocytes were considered as recipients. No differences in the rates of fusion (75.7, 77.7, 76.3 and 86.7%) and cleavage (80.1, 84.3, 77.8 and 74%) were detected among couplets reconstructed with FFC-40, FFC-65, AFC and ACC, respectively. Blastocyst formation rate of those oocytes reconstructed with FFC-40 was higher (18%; p < 0.001) than those reconstructed with FFC-65 (13%) and AFC (10.9) and comparable with those reconstructed with ACC (17.5%). When the effect of passage number was analysed within groups (FFC-40, FFC-65 and AFC) there were no significant differences in fusion, cleavage and blastocyst rates between reconstructed oocytes. The present study demonstrates that the fetal and adult fibroblasts as well as fresh cumulus cells are comparable in their ability to attain cell fusion and embryonic cleavage. Moreover, the blastocyst formation rate is influenced by the age of the donor animal and the fresh cumulus cells have similar remodelling potential to that of fetal fibroblasts in term of blastocyst formation rate.

Keywords

EmbryoFibroblastNuclear transferOvine
Type
Research Article
Information
Zygote , Volume 18 , Issue 4 , November 2010 , pp. 331 - 338
Copyright
Copyright © Cambridge University Press 2010

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