Addition of l-ascorbic acid to culture and vitrification media of IVF porcine blastocysts improves survival and reduces HSPA1A levels of vitrified embryos
Miriam Castillo-Martín A C , Marc Yeste B , Albert Soler A , Roser Morató A and Sergi Bonet A
+ Author Affiliations
- Author Affiliations
A Biotechnology of Animal and Human Reproduction (TechnoSperm), Department of Biology, Institute of Food and Agricultural Technology, University of Girona, Campus Montilivi, E-17071 Girona, Spain.
B Unit of Animal Reproduction, Department of Animal Medicine and Surgery, Faculty of Veterinary Medicine, Autonomous University of Barcelona, E-08193 Bellaterra, Spain.
C Corresponding author. Email: miriam.castillo@udg.edu
Reproduction, Fertility and Development 27(7) 1115-1123 https://doi.org/10.1071/RD14078
Submitted: 21 January 2014 Accepted: 21 March 2014 Published: 17 April 2014
Abstract
The aim of the present study was to determine the effect of l-ascorbic acid on embryo quality and gene expression of porcine blastocysts after supplementations of in vitro culture medium and/or vitrification–warming media. Embryo quality, in terms of total cell number (TCN), DNA fragmentation and peroxide levels, together with the relative transcript abundance of BCL-2 associated X protein (BAX), BCL2-like 1 (BCL2L1), POU class 5 homeobox 1 (POU5F1) and heat shock protein 70 (HSPA1A), was analysed. In Experiment 1, gene expression and embryo quality of fresh blastocysts were evaluated after culture with or without l-ascorbic acid; no significant differences were observed between the groups. In Experiment 2, blastocysts cultured with or without l-ascorbic acid were vitrified using two different vitrification solutions, supplemented or not with l-ascorbic acid. Supplementation of culture and vitrification media significantly enhanced survival rates and reduced peroxide levels. No significant differences in TCN, DNA fragmentation and BAX, BCL2L1 and POU5F1 expression were found in vitrified blastocysts among experimental groups. Vitrification procedures increase HSPA1A transcript abundance, but this increase was significantly lower in embryos cultured and/or vitrified with l-ascorbic acid. Thus, supplementing culture and/or vitrification media with l-ascorbic acid enhances survival rates of porcine blastocysts, suggesting a relationship with HSPA1A expression.
Additional keywords: antioxidant, Cryotop, gene expression, peroxide levels.
References
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