Osmotic stress induced by sodium chloride, sucrose or trehalose improves cryotolerance and developmental competence of porcine oocytes.
ABSTRACT Exposure of porcine oocytes to increased concentrations of NaCl prior to manipulation has been reported not only to increase cryotolerance after vitrification, but also to improve developmental competence after somatic cell nuclear transfer (SCNT). In the present study we compared the effects of NaCl with those of concentrated solutions of two non-permeable osmotic agents, namely sucrose and trehalose, on the cryotolerance and developmental competence of porcine oocytes. In Experiment 1, porcine in vitro-matured cumulus-oocyte complexes (COCs; n = 1200) were exposed to 588 mOsmol NaCl, sucrose or trehalose solutions for 1 h, allowed to recover for a further 1 h, vitrified, warmed and subjected to parthenogenetic activation. Both Day 2 (where Day 0 is the day of activation) cleavage and Day 7 blastocyst rates were significantly increased after NaCl, sucrose and trehalose osmotic treatments compared with untreated controls (cleavage: 46 +/- 5%, 44 +/- 7%, 45 +/- 4% and 26 +/- 6%, respectively; expanded blastocyst rate: 6 +/- 1%, 6 +/- 2%, 7 +/- 2% and 1 +/- 1%, respectively). In Experiment 2, COCs (n = 2000) were treated with 588 mOsmol NaCl, sucrose or trehalose, then used as recipients for SCNT (Day 0). Cleavage rates on Day 1 did not differ between the NaCl-, sucrose-, trehalose-treated and the untreated control groups (92 +/- 3%, 95 +/- 3%, 92 +/- 2% and 94 +/- 2%, respectively), but blastocyst rates on Day 6 were higher in all treated groups compared with control (64 +/- 2%, 69 +/- 5%, 65 +/- 3% and 47 +/- 4%, respectively). Cell numbers of Day 6 blastocysts were higher in the control and NaCl-treated groups compared with the sucrose- and trehalose-treated groups. In conclusion, treatment of porcine oocytes with osmotic stress improved developmental competence after vitrification combined with parthenogenetic activation, as well as after SCNT.
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ABSTRACT: Objective To evaluate whether accidental contamination of follicular fluid with endometrioma content during oocytes retrieval affects IVF outcome. Study Design We set up a retrospective review of women undergoing IVF in our Infertility Unit during a four years period. Cases were 19 women with ovarian endometriomas who had an accidental aspiration of the endometrioma content during oocyte retrieval. Controls (n = 38) were the two subsequent age-matched women with endometriomas who underwent an uncomplicated oocyte retrieval. The main outcome was to evaluate the clinical pregnancy rate in cases and in controls. Results Accidental aspiration of endometrioma content occurred in 19 out of 314 women (6.1%, 95%CI: 3.8-9.1%). The number of embryos and top quality embryos was significantly higher in exposed women (3.1 ± 2.0 versus 1.7 ± 1.2 and 1.9 ± 1.5 versus 1.1 ± 1.1, respectively) but the chances of pregnancy were lower. The adjusted RRs of clinical pregnancy and live birth were 0.63 (95%CI: 0.49-0.87, p = 0.005) 0 and 0.60 (95%CI: 0.51-086, p = 0.003), respectively. Conclusion Accidental contamination of the follicular fluid with endometrioma content is an uncommon but possible event. This situation may affect IVF outcome but does not justify systematic surgical removal before the cycle.European Journal of Obstetrics & Gynecology and Reproductive Biology 10/2014; · 1.63 Impact Factor
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ABSTRACT: Handmade cloning (HMC) has been used to generate transgenic pigs for biomedical research. Recently, we found that parthenogenetic activation (PA) of porcine oocytes and improved HMC efficiency could be achieved by treatment with sublethal high hydrostatic pressure (HHP). However, the molecular mechanism underlying the effects of HHP treatment on embryonic development is poorly understood and so was investigated in the present study. Thus, in the present study, we undertook genome-wide gene expression analysis in HHP-treated and untreated oocytes, as well as in 4-cell and blastocyst stage embryos derived by PA or HMC. Hierarchical clustering depicted stage-specific genomic expression profiling. At the 4-cell and blastocyst stages, 103 and 163 transcripts were differentially expressed between the HMC and PA embryos, respectively (P<0.05). These transcripts are predominantly involved in regulating cellular differentiation, gene expression and cell-to-cell signalling. We found that 44 transcripts were altered by HHP treatment, with most exhibiting lower expression in HHP-treated oocytes. Genes involved in embryonic development were prominent among the transcripts affected by HHP. Two of these genes (INHBB and ME3) were further validated by quantitative reverse transcription-polymerase chain reaction. We also observed that HHP treatment activated expression of the imprinting gene DLX5 in 4-cell PA embryos. In conclusion, our genomic expression profiling data suggest that HHP alters the RNA constitution in porcine oocytes and affects the expression of imprinting genes during embryonic development.Reproduction Fertility and Development 03/2014; 26(3):469-84. · 2.58 Impact Factor
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ABSTRACT: The aim of this study was to determine the effects of trans-10, cis-12 conjugated linoleic acid (t10c12 CLA) supplementation on oocyte maturation and embryo development in pigs. Compared with the control, supplementation of 50 µM t10c12 CLA to in vitro maturation (IVM) medium significantly increased the proportion of oocytes at the metaphase II (MII) stage and subsequent parthenogenetic embryo development in terms of cleavage rate, blastocyst formation rate, and cell numbers in blastocysts. The t10c12 CLA-treated oocytes resumed meiotic maturation and progressed to the MII stage significantly faster than those of control. The expression of phosphorylated mitogen-activated protein kinase 3/1 (p-MAPK3/1) and cyclooxygenase-2 (COX2) in cumulus oocyte complexes (COCs) at 5, 10, and 22 h of IVM were significantly increased in the t10c12 CLA-treatment group. The level of p-MAPK3/1 in t10c12 CLA-treated MII oocytes was also higher (P <0.05) than that of control. Moreover, t10c12 CLA supplementation partially overcame the negative effects of U0126 on cumulus expansion and nuclear maturation, and completely recovered COX2 protein levels in the presence of U0126. Treatment of COCs with NS398 also significantly suppressed cumulus expansion and nuclear maturation, which was overcome by t10c12 CLA. Yet, this simulatory effect of t10c12 CLA was blocked in the presence of both U0126 and NS398. The t10c12 CLA treatment significantly reduced reactive oxygen species level and increased glutathione concentrations in MII oocyte. In conclusion, supplementation of t10c12 CLA during porcine oocyte maturation exerts its beneficial effects on nuclear and cytoplasmic maturation, which contributes to enhancing subsequent embryo development. Mol. Reprod. Dev. © 2013 Wiley Periodicals, Inc.Molecular Reproduction and Development 10/2013; · 2.68 Impact Factor