Chatzimeletiou K.,Section of Reproductive Medicine |
Morrison E.E.,University of Leeds |
Panagiotidis Y.,Iakentro Advanced Medical Center |
Vanderzwalmen P.,Ivf Center Prof Zech |
And 4 more authors.
Human Reproduction | Year: 2012
BACKGROUND: Vitrification of human blastocysts is being used increasingly to cryopreserve supernumerary embryos following IVF. In this study, we investigate the effects of aseptic vitrification on the cytoskeleton and development of human blastocysts, by analysing survival rates and spindle and chromosome configurations by fluorescence and confocal laser scanning microscopy. METHODS: A total of 55 fresh blastocysts and 55 day 5 dimethylsulphoxide/ethylene glycol vitrified blastocysts, which were allowed to remain in culture for 24 h post-warming, were rapidly fixed in ice cold methanol, and immunostained with an a-tubulin antibody to visualize microtubules in combination with antibodies against acetylated tubulin (to visualize spindles, poles and mid bodies), gamma tubulin (to identify spindle poles) and 4(6-diamidino-2-phenylindole) to visualize DNA. RESULTS: In total, 213 spindles were analysed in the control (fresh) group of which 183/213 (85.9) were normal, 20/213 (9.4) were abnormally shaped, 9/213 (4.2) were multipolar and 1/213 (0.5) was monopolar. A total of 175 spindles were analysed in the vitrified group, of which 120/175 (68.6) were normal, 39/175 (22.3) were abnormally shaped, 10/175 (5.7) were multipolar and 6/175 (3.4) were monopolar. The incidence of multipolar spindles was similar in the two groups, but the level of abnormally shaped spindles, often associated with chromosome lagging, or congression failure, was significantly higher in the vitrified group compared with the fresh group (P< 0.05). CONCLUSIONS: The high survival rate following thawing and the large proportion of normal spindle/chromosome configurations suggests that vitrification at the blastocyst stage on Day 5 does not adversely affect the development of human embryos and the ability of spindles to form and continue normal cell divisions. However, there was a significantly higher incidence of abnormal spindles in the vitrified group compared with the fresh group, notably of spindles with a focused and an unfocused pole as well as chromosome bridging and disorganized middle spindle fibres at telophase. Further investigation is warranted to elucidate the mitotic stages that are more vulnerable to damage during vitrification, the fate of the abnormal spindles and any potential effects that may be reflected on the chromosomal constitution of the developing blastocysts. © The Author 2011.
Panagiotidis Y.,Iakentro Advanced Medical Center |
Panagiotidis Y.,Democritus University of Thrace |
Vanderzwalmen P.,Center Hospitalier Inter regional Cavell |
Vanderzwalmen P.,Ivf Centers Prof Zech |
And 10 more authors.
Reproductive BioMedicine Online | Year: 2013
The use of open carriers for embryo vitrification has raised safety concerns and therefore vitrification in closed systems has been proposed. However, the drop in the cooling rate emerges as a major drawback. The objective of the present study was to compare the efficiency of vitrification in open versus closed conditions. Blastocysts were randomly allocated either to open ultra-rapid vitrification (group I) or closed aseptic vitrification (group II). In group I, blastocysts were exposed to two solutions of ethylene glycol/dimethylsulphoxide (10%/10% and 20%/20%), while in group II, blastocysts were pretreated with a solution of lower concentration (5%/5%). A total of 208 and 224 vitrification-warming cycles were performed for groups I and II, respectively. Both groups were equal in terms of maternal age, sperm parameters and number and quality of blastocysts vitrified, warmed and transferred per cycle. Importantly, there was no significant difference between the groups in the analysed outcomes; embryo survival rate (84.1% versus 82.1%), clinical pregnancy rate (45.9% versus 42.4%), implantation rate (25.6% versus 24.5%), cycle cancellation rate (6.7% versus 8.5%) and live birth rate (41.2% versus 41.0%). These data suggest that ultra-rapid vitrification may be replaced by aseptic vitrification without affecting clinical efficiency. © 2013, Reproductive Healthcare Ltd. Published by Elsevier Ltd. All rights reserved.
Margioula-Siarkou C.,Aristotle University of Thessaloniki |
Margioula-Siarkou C.,Iakentro Advanced Medical Center |
Kalogiannidis I.,Aristotle University of Thessaloniki |
Kalogiannidis I.,Iakentro Advanced Medical Center |
And 9 more authors.
International Journal of Preventive Medicine | Year: 2015
Objective: To examine vertical transmission rates of Cytomegalovirus, Toxoplasma Gondii and Rubella infections according to amniotic fluid PCR analysis. Methods: A retrospective analysis of mid-trimester amniocenteses performed in in pregnancies with diagnosed maternal infection by Cytomegavirus (CMV), Rubella or Toxoplasma gondii during 1994-2008 was performed. Vertical transmission rates were observed according to the presence of the infectious agent’s DNA in the amniotic fluid. A univariate regression model was also performed to investigate possible correlations between transmission and epidemiological parameters. Results: Overall, 7033 amniocenteses were performed during study’s period, of which 166 (2.4%) with the indication of maternal infection by CMV, Rubella or Toxoplasma. Mean maternal age was 27.4 ± 2.5 years and the mean gestational age at amniocentesis was 18.7 ± 2.5 weeks. Vertical transmission was observed in 21 cases (12.7%). Transmission rate was 17.3% in cases with infection from CMV, 9.5% from Toxoplasma gondii and 7.8% from Rubella (P =.05). Maternal age was the only parameter being significantly associated with increased risk for vertical transmission (P =.04). Conclusions: According to our results, overall vertical transmission rate marginally exceeds 10%. CMV infection is characterized by relatively higher transplacental transmission rate, while increased maternal age appears to be associated with a higher risk for vertical transmission. © 2015 Margioula-Siarkou C..