203 EFFECT OF FRUCTOSE ON THE DEVELOPMENT OF BOVINE SEXED EMBRYOS IN VITRO
S. A. Chaubal A , J. Xu A , X. Yang B and F. Du AA Evergen Biotechnologies Inc., Vernon, CT, USA;
B University of Connecticut, Storrs, CT, USA
Reproduction, Fertility and Development 21(1) 199-200 https://doi.org/10.1071/RDv21n1Ab203
Published: 9 December 2008
Abstract
It is important to optimize the condition of culture to promote the development of embryos fertilized by sex-sorted sperm. Fructose is a monosaccharide hexose that can be used as a substrate for glycolysis. Recently, there has been interest in exploring the embryotrophic properties of fructose during bovine embryo culture. The present study was undertaken to ascertain the effect of supplementation of fructose in culture on the development of sexed embryos in vitro. A total of 2310 slaughterhouse dairy oocytes, spread over 3 experimental replicates, were subjected to routine in vitro maturation (IVM) for 22 h. The maturation medium was M-199 with Earl’s salts, L-glutamine, 26.19 mm sodium bicarbonate, and 25 mm HEPES plus 10% fetal bovine serum. Fertilization of IVM oocytes was performed in Brackett and Oliphant’s medium by using frozen and sorted X-sperm from a single bull. Groups of 50 matured oocytes were added to a final volume of 100 μL of fertilization medium, with a final concentration of 0.3 × 106 spermatozoa mL–1. The oocytes were incubated with X-sperm for 6 h in 5% CO2 humidified air at 39°C. Presumptive zygotes were randomly allocated to culture either in a control medium (6 mg mL–1 of BSA CR1aa medium) or with 4.5 mm fructose added to CR1aa culture medium for 7 days in an atmosphere of 5% O2, 5% CO2, and 90% N2 at 39°C. Embryo cleavage, and development to the morulae and blastocyst stages were evaluated on Days 2, 5, and 7 of culture, respectively. Embryos were graded according to International Embryo Transfer Society standards. The total number of blastocysts recorded was the sum of grade C1 and C2 embryos. Data were analyzed by using the General Linear Model (SPSS 11.0, SPSS Inc., Chicago, IL, USA). As shown in Table 1, addition of fructose to the culture medium affected neither the rate of cleavage nor subsequent development to the morula stage or total blastocyst development. However, the proportion of C1-grade embryos of the total blastocysts was significantly (P < 0.05) improved. This study suggests that addition of fructose to the culture medium can promote embryo quality and yields that are suitable for embryo transfer.
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