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: Igor I KatkovPrinciples and Practice of Fertility Preservation, 1st edited by J Donnez, S S Kim, 07/2010: chapter 14: pages 177-199; Cambridge University Press.
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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
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ABSTRACT: Introduction the embryo biopsy aims to select genetically normal embryos. This selection occurs through pre‐ implantation genetic testing. It is expected the reduction of risk of genetic disorders and increase implantation rates in IVF. Objective to verify, through bibliographical revision, which embryo biopsy technique is considered more suitable for pre‐implantation genetic diagnosis. Method bibliographical research, in the form of literary review of scientific publications via networks, US National Library of Medicine (Pubmed), Latin‐American Literature and Caribbean Health Sciences (Lilacs), Google Scholar and Virtual Health Library. Results and conclusion there are three ways to perform the biopsy on assisted human reproduction. The first one consists in removing the 1st and/or 2nd polar body (if there was fertilization). You can also perform the biopsy from the one blastomere of embryo cleavage stage or use 5‐10 trophoectoderm cells blastocyst. Usually the techniques used for diagnostic purpose are PCR, Fish, CGH array, SNP array and others. Nowadays it is believed that blastocyst biopsy is the best technique in order to maintain the embryonic implantation. This tendency is justified by the larger amount of genetic material available in an advanced stage of embryonic development. It is assumed that in this stage the incidence of mosaicism is reduced with the consequent increase in the effectiveness of genetic testing. Another important question is that the blastocyst biopsy cells are removed from the trophoectoderm while inbiopsy incleavage stage, the removal of oneblastomerecan impairembryonic development.Reprodução & Climatério. 01/2014;