Abstract
The 9-cis retinoic acid (9-cisRA) enhances early embryonic development in both in vitro and in vivo conditions. This experiment was conducted to evaluate the effect of supplementation of 9-cisRA in the in vitro maturation (IVM) medium on embryo development efficiency and embryo quality. For this purpose, immature cumulus oocyte complexes (COC) collected from slaughterhouse derived bovine ovaries were matured in three different IVM media (control group, DMSO group and DMSO+RA group). In the control group, base IVM medium were used without supplementation of 9-cisRA and DMSO. In the DMSO group, base IVM medium was supplemented with 0.5 μl DMSO per ml IVM medium without 9-cisRA. In DMSO+RA group, base medium was supplemented with 5 nm 9-cisRA dissolved in 0.5 μl DMSO. Data were analyzed using one way ANOVA method and means were compared using Duncan’s multiple range test. Results showed that, supplementation of 9-cisRA in the maturation medium has no effect on embryonic development uptocleavage stage. However, blastocyst development rates (P>0.01), total blastomere number (P> 0.01), number of apoptotic blastomere per blastocyst (P>0.05) and percent of apoptotic blastomere per blastocyst (P>0.05) were significantly influences by 9-cisRA. In conclusion, 9-cisRA may be supplemented into the maturation medium for increasing bovinein vitro blastocyst development efficiency and blastocyst quality.Asian J. Med. Biol. Res. December 2017, 3(4): 516-520
Highlights
The in vitro embryo production (IVP) and embryo transfer techniques have been adopted with traditional cattle breeding programme for multiplying high genetic merit cattle in many countries
Blastocyst development rates were calculated at day 8 as a proportion of the presumed zygote transferred into IVC-I medium
The stimulatory effects of retinoic acid on blastocyst development that are found in this study are consistent with previous reports (Deb et al, 2011; First and Parrish, 1987; Hidalgo et al, 2003)
Summary
The in vitro embryo production (IVP) and embryo transfer techniques have been adopted with traditional cattle breeding programme for multiplying high genetic merit cattle in many countries. Sperm were capacitated through incubation with 500 μL IVF medium (Tyrode’s lactate solution supplemented with 6 mg/mL BSA, 22 μg/mL sodium pyruvate, 100 IU/mL penicillin, and 0.1 mg/mL streptomycin) containing heparin sodium salt (20 μg/mL) for 15 min. The matured COC were co-cultured with capacitated spermatozoa for 18 to 20 h through placing them into IVF medium (500 to 700 μL per well of 4well dish). After IVF, the cumulus cells were removed by gentle pipetting into TL-HEPES and the denuded presumed zygotes were placed into a 100 μL droplet of CR1-aa medium (Rosenkrans et al, 1993), supplemented with 44 μg/mL Na-pyruvate, 14.6 μg/mL glutamine, 10 μL/mL penicillin/streptomycin, 3 mg/mL BSA and 310 μg/mL glutathione for 3 days (IVC-I).
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