FORMATION RATE AND CELLULAR COUNT OF BLASTOCYSTS OF RABBITS AS AFFECTED BY EPIDERMAL GROWTH FACTOR SUPPLEMENTATION TO CULTURE MEDIUM OF FRESH OR VITRIFIED MORULAE

Document Type : Original Article

Authors

1 Animal Production Research Institute, Agricultural Research Center, Egypt.

2 Department of Poultry Production, Faculty of Agriculture, Mansoura University, Mansoura, Egypt.

3 Department of Poultry Production, Faculty of Agriculture, Damietta University, Damietta , Egypt.

Abstract

The aim of this study was to evaluate the effect of addition of epidermal growth factor (EGF) to culture medium of fresh and vitrified rabbit embryos on formation rate into blastocysts/hatched blastocysts and quality of the embryos. Total of 16 mature NZW rabbit does were superovulated by PMSG and hCG. Embryos were recovered 64-66 h post-mating by flushing from the oviducts of the slaughtered animals. Compact morulae were used in this study. Embryos were divided into two groups, fresh (n=200) and intended for vitrification (n=185). Fresh (n=200) or vitrified embryos (n=150) were in vitro cultured. Either fresh or vitrified embryos were in vitro cultured in medium supplemented with EGF (n=100 and 75) as compared to control medium (n=100 and 75), respectively, under mineral oil in CO2 incubator at 38°C, 95% humidity and 5% CO2 in air. Embryos were assessed daily to record the developmental competence of embryos in term of formation rate of embryos at blastocyst from morula stage, from blastocyst to expanded blastocyst stage or from expanded to hatched blastocyst throughout 5 days as culture period. Total cell number and intrazonal diameter of blastocysts were determined. Results showed that formation rate of blastocyst (BLs), expanded BLs and hatched BLs was the highest (P<0.05) for fresh embryos cultured with EGF (86.00, 94.11 and 88.92%), moderate for fresh embryos without EGF (74.00, 81.11 and 75.23%) or those vitrified with EGF (70.62, 80.67 and 73.89%), and the lowest for vitrified embryos cultured without EGF (57.76, 70.00 and 56.76%). Adding EGF in culture medium increased (P<0.05) total cell number and intrazonal diameter of blastocysts produced from fresh (121.20/BLs and 129.2 µm) or vitrified (118.2/BLs and 124.6 µm) morulae as compared to fresh (109.0/BLs and 124.2 µm) or vitrified (105/BLs and 117.6 µm) morulae cultured with free media.
In conclusion, supplementation of culture medium with 10 ng/ml epidermal growth factor (EGF) had beneficial effects of the developmental competence of fresh or vitrified embryos at morula stage to reach hatched blastocyst stage with good quality.

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