Vitrification of biopsied embryos at cleavage, morula and blastocyst stage.

Pedieos IVF Center, Nicosia, Cyprus.
Reproductive biomedicine online (Impact Factor: 2.98). 10/2009; 19(4):526-31. DOI: 10.1016/j.rbmo.2009.05.009
Source: PubMed

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.

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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.
    01/2014; 28(3). DOI:10.1016/j.recli.2014.06.001
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    ABSTRACT: Preimplantation genetic diagnosis and/or screening (PGD/PGS) allow the assessment of the genetic health of an embryo before transferring it into the uterus. These techniques require the removal of cellular material (polar bodies, blastomere(s) or trophectoderm cells) in order to perform the proper genetic analysis. We report the implantation and live birth outcome of a vitrified-warmed blastocyst developed after triple biopsy and double vitrification procedures at oocyte, cleavage embryo and blastocyst stage. An infertile couple, with family history of β-thalassemia, searched for IVF procedure and PGD. First polar bodies biopsy with subsequent vitrification was uninformative due to meiotic crossing-over, so oocytes were inseminated after warming. Two embryos were obtained and blastomere biopsy was performed on day 3 with inconclusive results on their genetic status. Their culture resulted in one expanded blastocyst stage on day 7 that underwent trophectoderm biopsy and vitrification. This embryo showed to be normal. It was then warmed and transferred in an artificial cycle. Preconception genetic analysis by removal and analysis of the first polar body is technically possible, but the genetic information that we can obtain at this stage may be limited and the oocytes to be inseminated is not predictable. Compared to blastomere biopsy, trophectoderm biopsy has more diagnostic efficiency with respect to both chromosomal mosaicism and PCR accuracy, reducing the problems of amplification failure and allele drop out. Moreover, embryos biopsied at the cleavage stage seem to have lower implantation rate than biopsied blastocyst. This is the first case report of a live birth obtained from a three step biopsy and double vitrification procedures of a blastocyst. This case report seems also to suggest the harmlessness of all these procedures if carefully performed by a skilled biologist in an IVF lab with quality management system. Finally, our study highlight that blastocyst cryopreserved on day 7 have clinically important potential and embryos that not reach blastocyst stage on day 6 should not to be discharged because they may result in an ongoing pregnancy.
    SpringerPlus 01/2015; 4:22. DOI:10.1186/s40064-015-0788-y
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    ABSTRACT: Este artículo debe citarse como: García-Amador MI, Martínez-Armas R, Ruvalcaba-Castellón LA. Vitrificación de ovocitos y embriones. Rev Mex Reprod 2011;3(4):143-149. E n 1983 Trounson y col. 1 fueron los primeros en sumergir un ovocito directamente en nitrógeno líquido con aceptables tasas de supervivencia y fertilización, pero bajas de segmentación. En 1984, Zeilmarker y col. comunicaron el primer nacimiento luego de transferir embriones descongelados. 2 El primer nacido de un ovocito criopreservado fue reportado por Chen en 1986, quien utilizó la congela-ción lenta y dimetilsulfóxido (DMSO). 3 Esta técnica fue mejorando a través de los años como resultado de cambios en los componentes de los medios de cultivo utilizados: medios basados en colina, 4 reducidos en so-Artículo de revisión Revista Mexicana de Medicina de la Reproducción 2011;3(4):143-149 RESUMEN En 1983, Trounson y colaboradores fueron los primeros en sumergir un ovocito directamente en nitrógeno líquido con tasas acep-tables de supervivencia y fertilización, pero bajas de segmentación. En 1984, Zeilmarker y su grupo reportaron el primer nacimiento luego de transferir embriones descongelados. En este artículo se revisan los aspectos generales de esta técnica y los resultados de la vitrificación de ovocitos, cigotos y embriones. Palabras clave: vitrificación, criopreservación, vitrificación de ovocitos, vitrificación de embriones.