Open pulled straw vitrification and slow freezing of sheep IVF embryos using different cryoprotectants
M. H. Bhat A G , V. Sharma B , F. A. Khan A , N. A. Naykoo A , S. H. Yaqoob C , G. Vajta D , H. M. Khan E , M. R. Fazili F , N. A. Ganai A and R. A. Shah A
+ Author Affiliations
- Author Affiliations
A Division of Biotechnology, Faculty of Veterinary Science, Sher-e-Kashmir University of Agricultural Sciences and Technology, Shuhama, Srinagar, 190006, Jammu and Kashmir, India.
B Department of Bio-Science & Biotechnology, Banasthali University, Rajasthan-304 022, India.
C Department of Animal production, College of Food and Agricultural Sciences, King Saud University, Riyadh 11451, Kingdom of Saudi Arabia.
D IRIS Central Queensland University, Rockhampton, Qld 4702, Australia.
E Mountain Research Centre for Sheep and Goat, Faculty of Veterinary Sciences, Sher-e-Kashmir University of Agricultural Sciences and Technology, Shuhama, Srinagar, 190006, Jammu and Kashmir, India.
F Teaching Veterinary Clinical Services Complex, Faculty of Veterinary Sciences, Sher-e-Kashmir University of Agricultural Sciences and Technology, Shuhama, Srinagar, 190006, Jammu and Kashmir, India.
G Corresponding author. Email: maajidhassan@yahoo.com
Reproduction, Fertility and Development 27(8) 1175-1180 https://doi.org/10.1071/RD14024
Submitted: 23 January 2014 Accepted: 18 April 2014 Published: 29 May 2014
Abstract
The aim of the present study was to evaluate the post-thaw survival and hatching rates of sheep blastocysts using different cryoprotectants. In Experiment 1, Day 6 sheep embryos were cryopreserved by a slow freezing protocol using 10% ethylene glycol (EG), 10% dimethyl sulfoxide (DMSO) or a mixture of 5% EG and 5% DMSO. Hatching rates were higher in the 10% EG group than in the 10% DMSO or EG + DMSO groups (30% vs 18% and 20%, respectively). In Experiment 2, embryos were cryopreserved by open pulled straw (OPS) vitrification using either 33% EG, 33% DMSO or a mixture of 16.5% EG + 16.5% DMSO. Re-expansion and hatching rates in the EG + DMSO group (79.16% and 52.74%, respectively) were higher than those in the EG group (64.28% and 30.02%, respectively), whereas the outcomes for the DMSO group were the lowest (45.18% and 8.6%, respectively). In Experiment 3, embryos were cryopreserved by OPS vitrification using either 40% EG, 40% DMSO or a mixture of 20% EG + 20% DMSO. Re-expansion and hatching rates were highest in the EG group than in the EG + DMSO and DMSO groups (92.16% vs 76.30% and 55.84% re-expansion, respectively; and 65.78% vs 45.55% and 14.46% hatching, respectively). In conclusion, OPS vitrification was found to be more efficient for cryopreservation of in vitro-developed sheep embryos than traditional freezing.
Additional keywords: blastocyst, in vitro production, vitrification.
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