Somatic cell nuclear transfer in buffalos: effect of the fusion and activation protocols and embryo culture system on preimplantation embryo development
Liz Simon A C , C. Veerapandian A , S. Balasubramanian B and A. Subramanian AA Department of Animal Reproduction, Gynecology and Obstetrics, Madras Veterinary College, Chennai 600010, India.
B Department of Clinics, Madras Veterinary College, Chennai 600010, India.
C Corresponding author. Email: lizfm2001@yahoo.co.in
Reproduction, Fertility and Development 18(4) 439-445 https://doi.org/10.1071/RD05079
Submitted: 7 July 2005 Accepted: 15 January 2006 Published: 22 March 2006
Abstract
The present study was conducted primarily to evaluate several factors that affect the nuclear transfer programme in water buffalos, in which relatively few studies have been performed. Embryos reconstructed with quiescent fetal fibroblasts and metaphase II cytoplasts were matured for 24 h, and activation was found to be comparatively better than in those matured for 30 h. A significantly higher proportion of embryos fused (52.0 ± 1.9) and cleaved (51.2 ± 1.7) when the couplets were fused 4–6 h before activation than when fused and activated simultaneously (46.5 ± 1.6 and 44.5 ± 2.0, respectively). Development of nuclear transfer embryos to the blastocyst stage (4.8 ± 2.2) was supported by a commercially available sequential medium, and cleavage (76.5 ± 2.8) was significantly higher in this medium compared with cleavage in TCM-199 with oviduct epithelial cell coculture (45.6 ± 1.5) and synthetic oviduct fluid (21.8 ± 6.6). Of the 16 cloned embryos transferred, none resulted in pregnancy. The present study demonstrates that optimal numbers of cloned buffalo blastocysts can be obtained from oocytes matured for 24 h, fused 3–4 h before activation and cultured in a commercially available sequential media (G1/G2), thus providing further information to enable successful nuclear transfer in buffalos.
Extra keyword: in vitro culture.
Acknowledgments
This study was supported by the Tamilnadu Veterinary and Animal Sciences University, Chennai, India and by a Senior Research Fellowship (to LS) from the Indian Council of Agricultural Research, Government of India.
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