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Deng A.,Vivere Health | Wang W.-H.,Houston Fertility Institute | Wang W.-H.,Center for Reproductive Medicine
Molecular Cytogenetics | Year: 2015

Background: Increased embryo implantation rates were reported after transfer of euploid embryos selected by preimplantation genetic screening (PGS). Egg cryopreservation by vitrification has become one of the most important assisted human reproduction technologies. Although reports indicate that development and implantation of human embryos derived from frozen donor eggs are comparative to fresh eggs, it is still unknown whether egg vitrification increases chromosomal abnormalities in eggs, which in turn causes formation of embryonic aneuploidy. Therefore, in this study, we evaluated the aneuploidy formation in the blastocysts derived from frozen donor eggs and also evaluated the efficiency of egg vitrification as an advanced technology for egg cryopreservation. Results: In this study, donated human eggs from young women were cryopreserved by vitrification and PGS was performed in the resulted blastocysts by DNA microarray. A total of 764 frozen eggs from 75 egg thawing cycles were warmed and 38 blastocysts were biopsied for PGS before embryo transfer. A 97.1% of egg survival rate was obtained and 59.1% of embryos developed to blastocyst stage. After biopsy and PGS, it was found that 84.2% of blastocysts were euploid and 15.8% were aneuploid. Aneuploidy rates varied among donors. Transfers of blastocysts without PGS resulted in higher clinical pregnancy and implantation rates as compared with transfer of blastocysts with PGS. Conclusions: Although the overall aneuploidy rate was low in the blastocysts derived from frozen donor eggs, high aneuploidy rates were observed in the embryos resulting from some donated eggs. Clinical pregnancy rate was not improved by PGS of embryos resulting from donor eggs, indicating that PGS may not be necessary for embryos derived from donor eggs in most cases. © 2015 Deng and Wang; licensee BioMed Central.

Qiao J.,Peking University | Wang Z.-B.,CAS Institute of Zoology | Feng H.-L.,Cornell University | Miao Y.-L.,U.S. National Institutes of Health | And 9 more authors.
Molecular Aspects of Medicine | Year: 2014

It is well known that maternal ageing not only causes increased spontaneous abortion and reduced fertility, but it is also a high genetic disease risk. Although assisted reproductive technologies (ARTs) have been widely used to treat infertility, the overall success is still low. The main reasons for age-related changes include reduced follicle number, compromised oocyte quality especially aneuploidy, altered reproductive endocrinology, and increased reproductive tract defect. Various approaches for improving or treating infertility in aged women including controlled ovarian hyperstimulation with intrauterine insemination (IUI), IVF/ICSI-ET, ovarian reserve testing, preimplantation genetic diagnosis and screening (PGD/PGS), oocyte selection and donation, oocyte and ovary tissue cryopreservation before ageing, miscarriage prevention, and caloric restriction are summarized in this review. Future potential reproductive techniques for infertile older women including oocyte and zygote micromanipulations, derivation of oocytes from germ stem cells, ES cells, and iPS cells, as well as through bone marrow transplantation are discussed. © 2013 Elsevier Ltd. All rights reserved.

Su Y.,Center for Reproductive Medicine | Li J.-J.,Center for Reproductive Medicine | Wang C.,Vivere Houston Fertility Laboratory | Haddad G.,Houston Fertility Institute | And 3 more authors.
Reproductive Biology and Endocrinology | Year: 2016

Background: Human embryos produced by in vitro fertilization (IVF) are usually cultured to day 6 after insemination, and good quality of embryos should develop to blastocyst stage at days 5 and 6. However, some embryos develop slowly, thus they may form blastocysts on day 7. Most IVF laboratories culture embryos to day 6 and then discard retarded embryos that do not develop to blastocyst stage. It has been reported that transfer of day 7 blastocysts can yield pregnancy although the rates were very low. In the present study, we evaluated the prevalence of aneuploidy in day 7 human blastocysts and also assessed embryo implantation after transfer of normal euploid blastocysts developed on day 7. Methods: Day 7 blastocysts were biopsied and screened for aneuploidy. Embryo implantation was assessed by transferring of euploid blastocysts. Results: A total of 1966 blastocysts from 367 IVF cycles were biopsied and screened for aneuploidy. It was found that 81.5 % of the patients had days 5 and 6 blastocysts and 18.5 % (68) patients had blastocysts developed on day 7, including 15.3 % had days 5-7 blastocysts and 3.3 % had only day 7 blastocysts. A total of 151 day 7 blastocysts, which accounted for 7.7 % of total blastocysts, were analyzed. It was found that 36.7 % of the blastocysts were euploid and 63.3 % had abnormal chromosomes, including aneuploidy and euploid with partial chromosome deletion. The aneuploidy rate was also maternal age dependent and was as high as 91.7 % in patients who were ≥40 years old. During the study period, transfer of day 7 euploid blastocysts in 15 patients resulted in 2 healthy live births. Conclusion(s): Aneuploidy rates in day 7 human blastocysts produced by IVF are very high. However, good euploid blastocysts have potential to implant and transfer of day 7 euploid blastocysts can result in healthy live birth. It is suggested that day 7 blastocyst culture may be necessary in patients who need aneuploidy screening. © 2016 Su et al.

Li L.,Lanzhou University | Zhang X.,Lanzhou University | Wang W.,Houston Fertility Institute
Zygote | Year: 2014

Summary Fluorescence in situ hybridization (FISH) is a cytogenetic technology used to detect chromosomal abnormalities in preimplantation human embryos. However, its efficiency is not stable due to improper sample preparation. The present study was designed to modify the current sample preparation technique and then to evaluate its efficiency in human preimplantation genetic diagnosis (PGD). Day 3 cleavage embryos as well as day 5 and 6 blastocysts were biopsied by mechanical aspiration method. In the present study, two methods were used for sample preparation of the biopsied cells. Method I was the traditional method, in which each blastomere was placed in a hypotonic solution for 5 min and then fixed on glass slides. The slides were kept at room temperature before the FISH procedures. Method II was a modified method, in which all blastomeres were placed individually in hypotonic solution drops covered by oil for at least 5 min and then fixed on slides with 0.1% Tween/HCl. After fixation, the slides were kept at -20°C for at least 30 min before the FISH procedures. The two methods were compared in terms of time consumption and proportions of blastomeres with FISH signals. In total, 329 blastomeres from day 3 embryos were fixed by Method I with an average fixation time of 8-10 min for each blastomere. By contrast, with Method II, 362 blastomeres were fixed and the average time was 3-4 min for each blastomere. After FISH, more nuclei had signals with Method II (97.2%) than with Method I (86.9%). All cells that were biopsied from blastocysts and prepared with Method II had FISH signals. However, Method I was not suitable for the fixation of multiple cells biopsied from blastocysts as cells were not traceable during the fixation. The present study indicates that proper sample preparation is critical for obtaining FISH signals in cells biopsied from preimplantation human embryos; hence these modifications can increase the efficiency of human PGD. © Cambridge University Press 2013.

Li L.,Lanzhou University | Zhang X.,Lanzhou University | Zhao L.,Lanzhou University | Xia X.,Methodist Hospital Research Institute | Wang W.,Houston Fertility Institute
Molecular Reproduction and Development | Year: 2012

Vitrification is a novel cryopreservation method for mammalian blastocysts. This study was designed to compare different vitrification methods and slow freezing for their effects on survival rate and DNA integrity in mouse and human blastocysts. In Experiment 1, embryo survival and DNA integrity were compared between mouse blastocysts with collapsed and non-collapsed blastoceles. In Experiment 2, embryo survival and DNA integrity were compared between vitrified and slow-frozen mouse blastocysts. In Experiment 3, embryo survival and DNA integrity were compared between vitrified and slow-frozen human blastocysts. Fresh blastocysts were used as controls in all experiments. Higher (P<0.05) blastocyst survival rates were obtained in mouse blastocysts vitrified with collapsed versus intact blastoceles, although DNA-integrity indices in the surviving blastocysts were the same among vitrified and fresh blastocysts. More mouse blastocysts (P<0.05) survived after vitrification (100%) as compared to slow freezing (82.5%). DNA-integrity indices examined in the surviving blastocysts were also higher (P<0.001) in fresh (93.6%) and vitrified/warmed (93.7%) blastocysts than in slow-frozen/thawed (75.8%) ones. More human blastocysts survived with a higher DNA-integrity index after vitrification/warming than after slow freezing/thawing. These results indicate that higher survival rates can be obtained by vitrification of blastocele-collapsed blastocysts, and that vitrification causes less cell apoptosis in both mouse and human blastocysts compared to slow freezing. Vitrification of blastocysts after blastocele collapse by single laser pulse supports a higher survival rate and less DNA apoptosis, suggesting that laser blastocele collapse is a safe procedure for blastocyst vitrification. © 2011 Wiley Periodicals, Inc.

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