Publications by authors named "Anne-Sophie Vannin"

9 Publications

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Outcomes of immature oocytes collected from ovarian tissue for cryopreservation in adult and prepubertal patients.

Reprod Biomed Online 2017 Jun 20;34(6):575-582. Epub 2017 Mar 20.

Research Laboratory on Human Reproduction, Campus Erasme, Université Libre de Bruxelles (ULB), Belgium; Fertility Clinic, Department of Obstetrics and Gynecology, CUB-Erasme Hospital, Université Libre de Bruxelles (ULB), Belgium.

The efficiency of oocyte in-vitro maturation (IVM) and vitrification procedures after ex-vivo collection from ovarian tissue were assessed according to patient age, number of retrieved oocytes and tissue transport conditions. The combined procedure was performed in 136 patients: 130 adults (mean 27.6 ± 5.6 years) and six prepubertal girls (mean 8.7 ± 2.3 years). A higher mean number of oocytes were collected in girls compared with adults (11.5 ± 8.0 versus 3.8 ± 4.2, respectively, P < 0.001) but the percentage of degenerated oocytes was significantly higher in girls (35.5% versus 17.1%, respectively, P < 0.001). IVM rates were significantly lower in prepubertal than postpubertal population (10.3% versus 28.1%, P = 0.002). In adults, a negative correlation was observed between number of retrieved oocytes and age (P = 0.002; r = -0.271); the correlation was positive between anti-Müllerian hormone (AMH) and number of collected oocytes (P = 0.002; r = 0.264). IVM rates were not correlated with AMH levels (r = 0.06) or age (r = -0.033). At present, nine oocytes and one embryo have been warmed in four patients and one biochemical pregnancy obtained. This suggests the combined procedure could be an additional option for fertility preservation.
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http://dx.doi.org/10.1016/j.rbmo.2017.03.007DOI Listing
June 2017

A randomized controlled trial comparing two vitrification methods versus slow-freezing for cryopreservation of human cleavage stage embryos.

J Assist Reprod Genet 2014 Feb 8;31(2):241-7. Epub 2013 Dec 8.

Research Laboratory on Human Reproduction, Faculty of Medicine, Université Libre de Bruxelles, Campus Erasme, Bruxelles, Belgium,

Purpose: To compare two different vitrification methods to slow freezing method for cryopreservation of human cleavage stage embryos.

Design: Prospective randomised trial.

Setting: University assisted reproduction centre.

Patient(s): 568 patients (mean age 33.4 ± 5.2) from April 2009 to April 2011.

Methods: 1798 supernumerary good-quality cleavage stage embryos in 645 IVF cycles intended to be cryopreserved were randomly allocated to three groups: slow freezing, vitrification with the Irvine® method, vitrification with the Vitrolife® method.

Main Outcome Measure(s): Embryo survival and cleavage rates, implantation rate.

Results: A total of 1055 embryos were warmed, 836 (79.2%) survived and 676 were finally transferred (64.1%). Post-warming embryos survival rate was significantly higher after vitrification (Irvine: 89.4%; Vitrolife: 87.6%) than after slow freezing (63.8%) (p < 0.001). No differences in survival rates were observed between the two vitrification methods, but a significant higher cleavage rate was observed using Irvine compared to Vitrolife method (p < 0.05). Implantation rate (IR) per embryo replaced and per embryo warmed were respectively 15.8% (41/259) and 12.4% (41/330) for Irvine, 17.0% (40/235) and 12.1% (40/330) for Vitrolife, 21.4% (39/182) and 9.9% (39/395) for slow-freezing (NS).

Conclusions: Both vitrification methods (Irvine and Vitrolife) are more efficient than slow freezing for cryopreservation of human cleavage stage embryos in terms of post-warming survival rate. No significant difference in the implantation rate was observed between the three cryopreservation methods.
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http://dx.doi.org/10.1007/s10815-013-0145-4DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3933602PMC
February 2014

Development and evaluation of single sperm washing for risk reduction in artificial reproductive technology (ART) for extreme oligospermic HIV positive patients.

Curr HIV Res 2008 Sep;6(5):461-5

Human Reproduction Research Laboratory, Université Libre de Bruxelles, Bruxelles, Belgium.

The serodiscordant couples, where the male is HIV-positive, are treated in fertility clinics, using the sperm washing technique by gradient centrifugation. This protocol cannot be carried out in oligo-azoospermic patients, where spermatozoa retrieval from the epididymis and testis must be performed. We developed a single sperm washing technique, where the spermatozoa, after the retrieval, are washed with the aid of a micromanipulator, to obtain virus decontamination and then used for the intracytoplasmic sperm injection (ICSI). The experiment was performed by using sperm samples containing three different viral loads. After one hour of incubation, spermatozoa were taken one by one from the HIV loaded drop and washed in four different microdrops. Before each passage into the next washing drop, the pipette was emptied in a first waste drop and then loaded with new washing medium from a second separate loading drop. After transferring of 10 spermatozoa in these four successive drops, the washing medium and the virus-loaded drops were tested for the HIV RNA presence by the nested RT-PCR technique. The presence of the virus was detected in the waste drop of all three viral loads. The four washing microdrops were each time negative for the presence of HIV-1 RNA, tested by the nested RT-PCR technique. The results show that by rinsing the spermatozoa four times, we are able to diminish the viral load to an undetectable level. Our data demonstrate that single sperm washing can be performed in the cases of extreme male sterility in HIV-positive men. From now on the couples, where the male is oligoazoospermic and HIV positive, could be included in our ICSI program, respecting the usual viral safety level of the ART techniques for the embryo.
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http://dx.doi.org/10.2174/157016208785861168DOI Listing
September 2008

Impaired ovarian stimulation during in vitro fertilization in women who are seropositive for hepatitis C virus and seronegative for human immunodeficiency virus.

Fertil Steril 2007 Sep 22;88(3):607-11. Epub 2007 Feb 22.

Laboratory for Human Reproduction Research, Faculty of Medicine, Campus Erasme, Université Libre de Bruxelles, Brussels, Belgium.

Objective: To analyze the impact of seropositivity with hepatitis C virus (HCV) on in vitro fertilization (IVF) outcomes.

Design: Retrospective, case-controlled study.

Setting: Fertility clinic of academic hospital.

Patient(s): 42 IVF/intracytoplasmic sperm injection cycles in HCV-seropositive women and 84 matched control cycles.

Intervention(s): IVF/intracytoplasmic sperm injection treatment for infertility.

Main Outcome Measure(s): Ovarian response to stimulation, laboratory findings, and implantation and pregnancy rates.

Result(s): Absence of ovarian response was statistically significantly higher for HCV-seropositive women compared with controls (10/42 vs 5/84 cycles, respectively). For cycles with oocyte retrieval, HCV-seropositive women required more gonadotropin units compared with controls. The maximum estradiol levels and number of collected oocytes were similar, but HCV-seropositive women had statistically significantly fewer embryos available compared with controls. Embryo morphologic features, number of transferred embryos, and rates of implantation and pregnancy were similar for HCV-seropositive women and controls.

Conclusion(s): When compared with matched uninfected controls, HCV-seropositive women display a decreased ovarian response.
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http://dx.doi.org/10.1016/j.fertnstert.2006.11.177DOI Listing
September 2007

Impact of the assessment of early cleavage in a single embryo transfer policy.

Reprod Biomed Online 2006 Aug;13(2):255-60

Fertility Clinic Erasmus Hospital, Free University of Brussels, French Speaking, Route de Lennik 808, 1070 Brussels Belgium.

The policy of single embryo transfer (SET) adopted for women <36 years old since 1 July 2003, strongly calls for improvement of embryo selection. A total of 196 cycles in which SET was performed were randomly allocated to two groups. In the first group, early cleavage was assessed (ECA) 25 h after insemination. The embryo with the best score that cleaved early, if present, was selected for transfer. In the second group, early cleavage was not assessed (ECNA) and embryo selection was based solely on the embryo score. Ninety-eight cycles were allocated in the ECA and ECNA group respectively. Early cleavage occurred in 64% of cycles and 32.2% of embryos. Patient population and embryo morphology were similar between the two groups, and similar delivery rates were observed (27.6 versus 24.5% respectively in the ECA and ECNA groups). The assessment of early cleavage as additional parameter did not improve the delivery rate in the single embryo transfer policy.
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http://dx.doi.org/10.1016/s1472-6483(10)60623-2DOI Listing
August 2006

Development and evaluation of a qualitative reverse-transcriptase nested polymerase chain reaction protocol for same-day viral validation of human immunodeficiency virus type 1 ribonucleic acid in processed semen.

Fertil Steril 2006 Jul 6;86(1):121-8. Epub 2006 Jun 6.

Fertility Clinic, Department of Obstetrics and Gynaecology and Laboratory for Research on Human Reproduction, Campus Erasme, Universite Libre de Bruxelles (ULB), Brussels, Belgium.

Objective: To develop a method for same-day validation of processed semen in the setting of assisted reproductive techniques (ART) with patients who are seropositive for human immunodeficiency virus, type 1 (HIV-1).

Design: Laboratory experiments.

Setting: University hospital.

Patient(s): Volunteers who are HIV-1 seronegative and seropositive.

Intervention(s): Evaluation of the sensitivity of a reverse-transcriptase (RT)-nested polymerase chain reaction (PCR) in HIV-1 RNA-positive blood plasma, in artificially infected blood plasma and semen, and in 85 semen samples of 29 HIV-1-seropositive volunteers. Semen was submitted to gradient separation, followed by swim-up.

Main Outcome Measure(s): Qualitative detection of HIV-1 RNA in blood plasma and in different parts of semen preparation by using RT-nested PCR, PCR inhibition control by dilution of samples, and an internal control.

Result(s): The detection limit of our PCR was 20 HIV-1 RNA copies per milliliter. Among seropositive patients, RNA was detected in 25% of fresh semen, 36.5% of seminal plasma, 27.5% of gradient supernatants, and 7.1% of final preparations before the migration-sedimentation stage. Positive final preparations were observed in patients who had blood viral loads of >/=20,000 HIV-1 RNA copies per milliliter. Inhibition was present in 17.6% of seminal plasma and in 20% gradient supernatants and in 2 final preparations among 69 tested. Among 25 preparations tested after the migration-sedimentation stage, 2 were positive (1 patient; 70,000 HIV-1 RNA copies per milliliter).

Conclusion(s): The RT-nested PCR detects low viral load and allows the validation of semen preparations of HIV-1-seropositive patients for ART on the day of sampling. For this purpose, the validation is performed on spermatozoa that are obtained after gradient separation before swim-up. Inhibition of the PCR must be controlled by using an internal control that is well-designed to explore the detection limit of the method.
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http://dx.doi.org/10.1016/j.fertnstert.2005.12.021DOI Listing
July 2006

Medically assisted reproduction in the presence of chronic viral diseases.

Hum Reprod Update 2004 Mar-Apr;10(2):149-62

Department of Obstetrics and Gynaecology, IVF Centre of the Université Libre de Bruxelles.

Teams practising medically assisted reproduction techniques try to avoid viruses as much as possible. Attitudes towards chronic carriers of viruses are rapidly changing, especially for human immunodeficiency virus (HIV) patients. We focus our attention on the legitimacy of systematic screening before assisted reproductive techniques and the need for specialized approaches including an adapted laboratory for viral hazards as well as the need for a multidisciplinary team. Specificities of HIV, hepatitis C virus (HCV), hepatitis B virus (HBV) carriers and the hypothesis of a reduced fertility potential are discussed. Are male HIV carriers a new indication for assisted reproductive techniques in order to prevent virus transmission? It is largely proven that sperm gradient preparation techniques efficiently decrease viral loads and therefore have a protective effect on contamination risk during assisted reproductive techniques. Although a few thousand assisted reproductive technique cycles were performed in the world for this indication without contamination, it is still too early to demonstrate that this technology is fully safe. Two examples of contaminations during insemination are examined. Many questions remain unresolved, such as the lack of standardized techniques for semen preparation or virus detection or the relative merits of intrauterine insemination or ICSI to prevent HIV contamination during assisted reproductive techniques. The authors plead for well-structured, separate programmes of care linked to research objectives.
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http://dx.doi.org/10.1093/humupd/dmh013DOI Listing
November 2004

Similar delivery rates in a selected group of patients, for day 2 and day 5 embryos both cultured in sequential medium: a randomized study.

Hum Reprod 2003 Oct;18(10):2145-50

Fertility Clinic, Erasme Hospital, Free University of Brussels, French Speaking, 808, Route de Lennik, B-1070 Brussels, Belgium.

Background: The existence of a real benefit of blastocyst transfer is still a matter of debate. The aim of this study was to compare, in a prospective randomized trial, the outcome of day 2 and day 5 transfer of human embryos cultured in an 'in-house' sequential medium.

Methods: A total of 193 cycles from 171 patients with less than four previous IVF cycles, <39 years old and with four or more zygotes on day 1, were randomly allocated to day 2 (94 cycles) or day 5 (99 cycles) transfer. Zygotes were kept in fertilization medium until 18 h post-fertilization and then placed in a 'glucose-free' cleavage medium. Embryos allocated for day 5 transfer were placed in a blastocyst medium 66 h post-fertilization. Two or three embryos were replaced according to the morphology.

Results: A mean (+/- SEM) number of 2.1 +/- 0.4 and 1.9 +/- 0.3 embryos were replaced on day 2 and day 5 (P < 0.001) respectively. Delivery rates per transfer were 44.1 and 37.1% [P = not significant (NS)], implantation rates were 31.4 and 29.4% (NS) and multiple delivery rates 22 and 36% (NS) for day 2 and day 5 groups respectively. Ten patients (10.1%) had no blastocysts available for transfer.

Conclusions: No clear benefits were observed using blastocyst transfer for patients aged <39 years who had had less than four previous IVF cycle attempts.
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http://dx.doi.org/10.1093/humrep/deg394DOI Listing
October 2003

Incidence of chromosomal mosaicism in human embryos at different developmental stages analyzed by fluorescence in situ hybridization.

Genet Test 2003 ;7(2):85-95

Fertility Clinic, Department of Obstetrics and Gynaecology, and Laboratory of Research on Human Reproduction, Erasmus Hospital-ULB, Free University of Brussels, Belgium.

Chromosomal mosaicism has been reported in in vitro-cultured embryos at early cleavage stages, as well as in morulae and blastocysts. We have assessed the incidence and pattern of mosaicism during in vitro development of human embryos from early-cleavage stages to morula and blastocyst. Fifty spare embryos were fixed for fluorescence in situ hybridization (FISH) analysis for chromosomes X, Y, 13, 18, and 21 on days 2 or 3 (4- to 10-cell stage) (n = 16), on day 4 (morula stage) (n = 14), on day 5 (pre-expanded blastocyst) (n = 5), and the expanded blastocyst stages (n = 15). Blocked embryos (no cleavage observed within the last 24 hr) were not included. A total of 2367 cells were analyzed. Four early-cleavage stage embryos were found uniformly diploid; all of the others were mosaic for the chromosomes analyzed (mean diploid nuclei 48.3% +/- 28.7). All of the embryos at more advanced developmental stages, except one fully normal morula, had mosaic chromosome constitutions, with an increase in the percentage of diploid cells in morulae, pre-expanded, and expanded blastocysts, respectively (mean diploid nuclei 78.6% +/- 11.7, 66.0% +/- 20.8, 79.6% +/- 12.8), in comparison with earlier stages. Hypotheses about the origin of mosaicism and embryo regulation mechanisms will be discussed.
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http://dx.doi.org/10.1089/109065703322146768DOI Listing
April 2004
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