Vitrification of biopsied embryos at cleavage, morula and blastocyst stage.
ABSTRACT This study investigated the effect of vitrification on biopsied embryos at various developmental stages. After biopsy on day 3, embryos were vitrified at cleavage, morula and blastocyst stages using a commercially available kit. Nonbiopsied embryos were vitrified as controls. For day-3 cleavage embryo vitrification, embryos from abnormally fertilized oocytes were randomly allocated to the biopsy and control groups. For morula and blastocyst vitrification, the embryos used in the biopsy groups were obtained from aneuploidy or affected embryos diagnosed by preimplantation genetic diagnosis (PGD). After warming, survival, blastulation and development of embryos in different groups were compared. The survival rate after warming in the non-biopsied cleavage control group was significantly higher than in the biopsied cleavage group (92.0% versus 64.0%, P = 0.037). Most of the biopsied embryos were destroyed due to blastomeres escaping. At the morula stage, both biopsied and non-biopsied embryos had similar survival rates. However, a significantly higher survival rate (95.6%) was observed in the biopsied blastocyst group compared with the control group (81.3%, P = 0.035). Biopsied embryos vitrified at an advanced stage had as high survival rates as non-biopsied embryos. Vitrification at the blastocyst stage is a practical and efficient solution for embryo cryopreservation during PGD.
- SourceAvailable from: Miquel Solé[show abstract] [hide abstract]
ABSTRACT: Embryos diagnosed as abnormal in Preimplantation Genetic Diagnosis (PGD) cycles are useful for the establishment of human Embryonic Stem Cells (hESC) lines with genetic disorders. These lines can be helpful for drug screening and for the development of new treatments. Vitrification has proved to be an efficient method to preserve human blastocysts. One hundred and three abnormal or undiagnosed vitrified blastocysts from the PGD programme at Institut Universitari Dexeus were donated for human embryonic stem cell derivation. The overall survival rate after warming was 70.6 %. Our results showed better survival rates when blastocysts have not started the hatching process (initial/expanded 87.8 %, hatching 68.3 % and hatched 27.3 %). Thirty-five blastocysts and 12 partially surviving embryos were seeded. One hESC line with the multiple exostoses type 2 paternal mutation was obtained.Journal of Assisted Reproduction and Genetics 06/2012; · 1.82 Impact Factor
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ABSTRACT: To compare success rates of vitrified-warmed with fresh and frozen-thawed ETs Retrospective. Public fertility center. Cryopreserved- thawed/warmed ETs were included in this study. Fresh cycles, in which supernumerary embryos were cryopreserved, were set as the fresh control group. Supernumerary day 3 embryos were cryopreserved by slow-freezing or vitrification and transferred after thawing or warming. Comparison of two cryopreservation techniques with respect to post-thaw survival of embryos, implantation and pregnancy rates, neonatal outcome, and congenital birth defects. A total of 962 fresh, 151 freezing-thawed and 300 vitrified-warmed cycles were included in this study. The survival and intact cell rates in the vitrification group were significantly higher compared with those in the slow freezing group (88.5 % vs 74.5 % and 86.6 % vs 64.0 %). The implantation, clinical pregnancy and live birth rates of the vitrification group were similar to the fresh and significant higher than slow freezing group. There were no significant differences in mean gestational age, birth weight, stillbirth, birth defects and the prevalence of neonatal diseases among three groups. Vitrified-warmed ETs yield comparable outcomes with fresh ETs and is superior to frozen-thawed ETs regarding the survival rate and clinical outcomes.Journal of Assisted Reproduction and Genetics 06/2012; 29(9):883-9. · 1.82 Impact Factor
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ABSTRACT: To determine the survival and subsequent in vitro development of human cleavage stage embryos and hatched blastocysts following varying periods of short-term storage at 4 °C, using tripronucleated human embryos (TPN) as a model. TPN cleavage embryos and hatched blastocysts short-term stored at 4 °C for 0 h (control), 24 h and 48 h. The main outcome measures were: survival rates (SR) and in vitro developmental ability (blastocyst rate and blastocyst-re-expansion rate) in each of the groups after storage. Cleavage-stage TPN survived at comparable rates to controls, regardless of storage time (average: 97.3 %). The in vitro development of cleavage-stage TPN stored for 24 h was comparable to that of controls (average 64.7 %), but was significantly impaired when storage lasted 48-h (20.8 %). After artificial shrinkage, SR was comparable in 24-h-stored and non-stored hatched blastocysts (85.7 %; p > 0.05), but was significantly impaired in the 48-h-stored group (20.0 %). Following 24-h storage, the re-expansion rate of hatched blastocysts was similar to that of controls (average: 57.1 %; p > 0.05), but was higher than that of the 48-h-stored group (15.0 %; p < 0.05). TPN human cleavage embryos and blastocysts can be successfully stored short-term for up to 24 h at 4 °C without using cryoprotectants without any significant negative impact on survival or subsequent in vitro development.Journal of Assisted Reproduction and Genetics 07/2013; · 1.82 Impact Factor