Does the S phase have an impact on the accuracy of comparative genomic hybridization profiles in single fibroblasts and human blastomeres?

Ramos, Laia; Rey, Javier del; Daina, Gemma; Martínez-Passarell, Olga; Rius, Mariona; Tuñón, Dolores; Campillo, Mercedes; Benet, Jordi; Navarro, J.

doi:10.1016/j.fertnstert.2013.10.031

Cited by 12 publications

(10 citation statements)

References 37 publications

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“…The obtained size range of fragments and amount of DNA for each WGA system were concordant with previously published data. [ 23 , 33 , 34 ] As expected, the size range of MDA products has been shown to be larger than other WGA commercial methods ( Fig 2 ). At the same time, it has been seen that the DNA size range and amplification amount product was not affected by the addition of DMSO in GEH-MDA reaction.…”

Section: Discussionsupporting

confidence: 72%

Novel Double Factor PGT strategy analyzing blastocyst stage embryos in a single NGS procedure

et al. 2018

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In families at risk from monogenic diseases affected offspring, it is fundamental the development of a suitable Double Factor Preimplantation Genetic Testing (DF-PGT) method for both single-gene analysis and chromosome complement screening. Aneuploidy is not only a major issue in advanced-maternal-age patients and balanced translocation carriers, but also the aneuploidy rate is extremely high in patients undergoing in vitro fertilization (IVF), even in young donors. To adequate NGS technology to the DF-PGT strategy four different whole genome amplification systems (Sureplex, MALBAC, and two multiple displacement amplification systems-MDA) were tested using TruSight One panel on cell lines and blastocyst trophectoderm biopsies-TE. Embryo cytogenetic status was analyzed by Nexus software. Sureplex and MALBAC DNA products were considered not suitable for PGT diagnosis due to inconsistent and poor results on Trusight one (TSO) panel. Results obtained with both MDA based methods (GEH-MDA and RG-MDA) were appropriate for direct mutation detection by TSO NGS platform. Nevertheless, RG-MDA amplification products showed better coverage and lower ADO rates than GEH-MDA. The present work also demonstrates that the same TSO sequencing data is suitable not only for the direct mutation detection, but also for the indirect mutation detection by linkage analysis of informative SNPs. The present work also demonstrates that Nexus software is competent for the detection of CNV by using with TSO sequencing data from RG-MDA products, allowing for the whole cytogenetic characterization of the embryos. In conclusion, successfully development of an innovative and promising DF-PGT strategy using TSO-NGS technology in TE biopsies, performed in-house in a single laboratory experience, has been done in the present work. Additional studies should be performed before it could be used as a diagnostic alternative in order to validate this approach for the detection of chromosomal aneuploidies.

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Section: Discussionsupporting

confidence: 72%

Novel Double Factor PGT strategy analyzing blastocyst stage embryos in a single NGS procedure

et al. 2018

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“…Depending on the WGA system used (DOP-PCR or SurePlex), the comprehensive cytogenetic analysis performed (mCGH or BAC aCGH) and the type of data correction applied, variable correlations between the incidence of segmental imbalances and the blastomeres' replicative cell-stage have been detected [20],[21].…”

Section: Introductionmentioning

confidence: 99%

Oligonucleotide Arrays vs. Metaphase-Comparative Genomic Hybridisation and BAC Arrays for Single-Cell Analysis: First Applications to Preimplantation Genetic Diagnosis for Robertsonian Translocation Carriers

et al. 2014

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Comprehensive chromosome analysis techniques such as metaphase-Comparative Genomic Hybridisation (CGH) and array-CGH are available for single-cell analysis. However, while metaphase-CGH and BAC array-CGH have been widely used for Preimplantation Genetic Diagnosis, oligonucleotide array-CGH has not been used in an extensive way. A comparison between oligonucleotide array-CGH and metaphase-CGH has been performed analysing 15 single fibroblasts from aneuploid cell-lines and 18 single blastomeres from human cleavage-stage embryos. Afterwards, oligonucleotide array-CGH and BAC array-CGH were also compared analysing 16 single blastomeres from human cleavage-stage embryos. All three comprehensive analysis techniques provided broadly similar cytogenetic profiles; however, non-identical profiles appeared when extensive aneuploidies were present in a cell. Both array techniques provided an optimised analysis procedure and a higher resolution than metaphase-CGH. Moreover, oligonucleotide array-CGH was able to define extra segmental imbalances in 14.7% of the blastomeres and it better determined the specific unbalanced chromosome regions due to a higher resolution of the technique (≈20 kb). Applicability of oligonucleotide array-CGH for Preimplantation Genetic Diagnosis has been demonstrated in two cases of Robertsonian translocation carriers 45,XY,der(13;14)(q10;q10). Transfer of euploid embryos was performed in both cases and pregnancy was achieved by one of the couples. This is the first time that an oligonucleotide array-CGH approach has been successfully applied to Preimplantation Genetic Diagnosis for balanced chromosome rearrangement carriers.

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“…When analyzing a biopsy for PGS, the cellular stage of the cell may possibly play a role in the embryo diagnosis. In a recent paper by Ramos et al (21), they showed that a majority (73.3%) of segmental imbalances were due to chromosome instability during cell division.…”

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Current experience concerning mosaic embryos diagnosed during preimplantation genetic screening

Harton¹,

Cinnioğlu²,

Fiorentino³

2017

Fertility and Sterility

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The concept of embryos containing multiple cell lines (mosaicism) is not new, but much attention has been paid to this concept recently owing to recent advances in molecular techniques to analyze human embryos. Mosaicism in embryos has been known and reported for some time, originally in early cleavage-stage embryos diagnosed with the use of fluorescence in situ hybridization (FISH). However, the early data have come under attack owing to the limited ability of FISH to reliably detect the actual copy number count of chromosomes as well as potential ascertainment bias of those early studies, which were all performed on already analyzed embryos found to be aneuploid. More recent molecular techniques for analyzing embryos have allowed scientists to really begin to understand mosaic embryos, and to now transfer and follow this class of embryo. Indeed, it could be said that three classes of embryos now exist after preimplantation genetic screening: euploid, aneuploid, and mosaic aneuploid. This paper attempts to bring to light the latest data on mosaic embryos and to understand how clinicians and others will deal with this issue today and in the future. Finally, an attempt is made to look to other fields of genetics to understand how this important issue can be dealt with as a group much better than any one individual group may be able to.

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Does the S phase have an impact on the accuracy of comparative genomic hybridization profiles in single fibroblasts and human blastomeres?

Cited by 12 publications

References 37 publications

Novel Double Factor PGT strategy analyzing blastocyst stage embryos in a single NGS procedure

Novel Double Factor PGT strategy analyzing blastocyst stage embryos in a single NGS procedure

Oligonucleotide Arrays vs. Metaphase-Comparative Genomic Hybridisation and BAC Arrays for Single-Cell Analysis: First Applications to Preimplantation Genetic Diagnosis for Robertsonian Translocation Carriers

Current experience concerning mosaic embryos diagnosed during preimplantation genetic screening

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