Preimplantation genetic diagnosis and the biopsy technique: Important considerations

Greco, Ermanno; Fabozzi, Gemma; Ruberti, Alessandra; Zavaglia, Daniela; Minasi, Maria Giulia

doi:10.4236/arsci.2013.12002

Cited by 8 publications

(5 citation statements)

References 56 publications

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“…The number of cells to be biopsied in the cleavage stage embryo may be one of the main issues in PGD [Greco et al 2013]. Some studies reported that biopsy of more than one blastomere from a cleavage stage embryo can negatively affect further development, blastocyst formation, and subsequently clinical outcomes [De Vos et al 2009;Goossens et al 2008].…”

Section: Discussionmentioning

confidence: 99%

Does sperm DNA fragmentation affect the developmental potential and the incidence of apoptosis following blastomere biopsy?

Haghpanah

Salehi

Novin

et al. 2015

Systems Biology in Reproductive Medicine

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Common methods employed in assisted reproduction technology (ART) include intracytoplasmic sperm injection (ICSI) with an unspecified level of sperm DNA fragmentation (SDF) and preimplantation genetic diagnosis (PGD). The aim of this study was to investigate the impact of SDF on human preimplantation embryo development and the incidence of apoptosis following a single blastomere biopsy. Using sperm chromatin dispersion (SCD) to assess SDF, a total of 20 processed semen samples were categorized into two groups; group I: SDF ≤30% and group II: SDF >30%. After ICSI, fertilization, cleavage, and embryo quality score were assessed. A single blastomere was biopsied from day 3 embryos and development was monitored on day 4. The frequency of apoptosis in biopsied embryos was assayed by TUNEL and the level of BCL-2, BAX, hsa-mir-15a, and hsa-mir-16-1 were assessed by quantitative real-time polymerase chain reaction (qRT-PCR). SCD was found to be negatively correlated with sperm motility and normal form spermatozoa (p < 0.05). The rate of fertilization, cleavage, and embryo quality score were not significantly different between the two groups (all p > 0.05). SDF >30% had no negative effect on potential development and did not increase the proportion of apoptotic cells and the level of apoptosis-related genes and microRNAs (miRNAs) in group II vs. group I (p > 0.05). It appears that at the levels assessed paternal genome damage had little if any negative effect on preimplantaton embryo development and apoptosis following single blastomere biopsy. This may reflect the selection of morphologically normal sperm for ICSI and the repair capacity of the oocyte.

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Section: Discussionmentioning

confidence: 99%

Does sperm DNA fragmentation affect the developmental potential and the incidence of apoptosis following blastomere biopsy?

Haghpanah

Salehi

Novin

et al. 2015

Systems Biology in Reproductive Medicine

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“…This technique has been adapted for comprehensive molecular cytogenetic analysis of metaphase II oocytes and their polar bodies [ 25 ], cleavage stage embryos [ 17 , 26 ], and blastocysts [ 27 ]. In this latter case, biopsy of trophectoderm (TE) cells can have several advantages with respect to biopsy of blastomeres at day 3 of embryo development [ 28 ]. Because a larger number of cells can be biopsied from a blastocyst, it is expected that more accurate information can be obtained as compared to one-cell or a few-cell biopsy from cleavage stage embryos, thus reducing the risk of misdiagnosis of embryonic chromosomal mosaicism.…”

Section: Introductionmentioning

confidence: 99%

Comparative Genomic Hybridization Selection of Blastocysts for Repeated Implantation Failure Treatment: A Pilot Study

Greco

Bono²,

Ruberti

et al. 2014

BioMed Research International

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The aim of this study is to determine if the use of preimplantation genetic screening (PGS) by array comparative genomic hybridization (array CGH) and transfer of a single euploid blastocyst in patients with repeated implantation failure (RIF) can improve clinical results. Three patient groups are compared: 43 couples with RIF for whom embryos were selected by array CGH (group RIF-PGS), 33 couples with the same history for whom array CGH was not performed (group RIF NO PGS), and 45 good prognosis infertile couples with array CGH selected embryos (group NO RIF PGS). A single euploid blastocyst was transferred in groups RIF-PGS and NO RIF PGS. Array CGH was not performed in group RIF NO PGS in which 1-2 blastocysts were transferred. One monoembryonic sac with heartbeat was found in 28 patients of group RIF PGS and 31 patients of group NO RIF PGS showing similar clinical pregnancy and implantation rates (68.3% and 70.5%, resp.). In contrast, an embryonic sac with heartbeat was only detected in 7 (21.2%) patients of group RIF NO PGS. In conclusion, PGS by array CGH with single euploid blastocyst transfer appears to be a successful strategy for patients with multiple failed IVF attempts.

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“…Potential sources of genetic material are: first (1 PB) and second polar bodies (2 PB), day 3 embryo blastomeres, and blastocyst trophectoderm cells (Brezina et al 2013 ; Greco et al 2013 ). All these techniques require several laboratory manipulation procedures before embryo transfer: oocyte zona pellucida laser drilling, embryo biopsy, cryopreservation and thawing, that might affect embryo survival and its implantation potential (Scott et al 2013a ; Zhang et al 2009 ).…”

Section: Introductionmentioning

confidence: 99%

Successful implantation and live birth of a healthy boy after triple biopsy and double vitrification of oocyte-embryo-blastocyst

Greco

Biricik²,

Cotarelo

et al. 2015

SpringerPlus

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IntroductionPreimplantation 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.Case descriptionAn 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.Discussion and EvaluationPreconception 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.ConclusionsThis 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.

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Preimplantation genetic diagnosis and the biopsy technique: Important considerations

Cited by 8 publications

References 56 publications

Does sperm DNA fragmentation affect the developmental potential and the incidence of apoptosis following blastomere biopsy?

Does sperm DNA fragmentation affect the developmental potential and the incidence of apoptosis following blastomere biopsy?

Comparative Genomic Hybridization Selection of Blastocysts for Repeated Implantation Failure Treatment: A Pilot Study

Successful implantation and live birth of a healthy boy after triple biopsy and double vitrification of oocyte-embryo-blastocyst

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