Extracellular embryo genomic DNA and its potential for genotyping applications

Galluzzi, Luca; Palini, Simone; Stefani, Stefania; Andreoni, Francesca; Primiterra, Mariangela; Diotallevi, Aurora; Bulletti, Carlo; Magnani, Mauro

doi:10.4155/fso.15.62

Cited by 56 publications

(77 citation statements)

References 19 publications

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“…Efforts have been made to develop noninvasive approaches for PGS (21,22). Palini et al (23) reported the observation of the existence of DNA in the blastocoele fluid, Gianaroli et al (24) performed a pilot study on chromosome screening using blastocentesis, and Stigliani et al (25,26) observed genomic and mitochondria DNA contents in the culture medium, which were correlated with embryo quality.…”

mentioning

confidence: 99%

Noninvasive chromosome screening of human embryos by genome sequencing of embryo culture medium for in vitro fertilization

Fang

Chen

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

199

218

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Preimplantation genetic screening (PGS) is widely used to select in vitro-fertilized embryos free of chromosomal abnormalities and to improve the clinical outcome of in vitro fertilization (IVF). A disadvantage of PGS is that it requires biopsy of the preimplantation human embryo, which can limit the clinical applicability of PGS due to the invasiveness and complexity of the process. Here, we present and validate a noninvasive chromosome screening (NICS) method based on sequencing the genomic DNA secreted into the culture medium from the human blastocyst. By using multiple annealing and looping-based amplification cycles (MALBAC) for whole-genome amplification (WGA), we performed next-generation sequencing (NGS) on the spent culture medium used to culture human blastocysts (n = 42) and obtained the ploidy information of all 24 chromosomes. We validated these results by comparing each with their corresponding whole donated embryo and obtained a high correlation for identification of chromosomal abnormalities (sensitivity, 0.882, and specificity, 0.840). With this validated NICS method, we performed chromosome screening on IVF embryos from seven couples with balanced translocation, azoospermia, or recurrent pregnancy loss. Six of them achieved successful clinical pregnancies, and five have already achieved healthy live births thus far. The NICS method avoids the need for embryo biopsy and therefore substantially increases the safety of its use. The method has the potential of much wider chromosome screening applicability in clinical IVF, due to its high accuracy and noninvasiveness.

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mentioning

confidence: 99%

Noninvasive chromosome screening of human embryos by genome sequencing of embryo culture medium for in vitro fertilization

Fang

Chen

et al. 2016

Proc. Natl. Acad. Sci. U.S.A.

199

218

View full text Add to dashboard Cite

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“…Mitochondrial DNA in particular has been evaluated as marker of blastocyst potential and implantation outcome [59]. More recently, gDNA in Day 5/6 medium has been evaluated for PGD applications, opening the possibility to retrieve genetic information of medical interest from the embryo without invasive procedures [60,61].…”

Section: New Low-invasive Approachesmentioning

confidence: 99%

Pre-implantation genetic diagnosis and screening: now and the future

Palini

Stefani

Primiterra

et al. 2015

Gynecological Endocrinology

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Since 1989, the year of the first pre-implantation genetic diagnosis (PGD), many developments occurred both in assisted reproduction techniques and in molecular tools. While PGD is a well-established and documented application, pre-implantation genetic screening (PGS) for the detection of aneuploid embryos is still debated due to the presence of mosaicism in the embryo, but especially to the knowledge of the limits that label an embryo as healthy or as appropriate to the life. The aim of this review is to present the state-of-the-art in the field of PGD and PGS, illustrating its benefits and limitations, along with biopsy techniques and the use of new high-throughput technologies.

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“…[66][67][68][69] Recently, cell-free DNA has also been detected in blastocyst-spent culture media but no agreement on concordance rate with the genetic status of embryos has been found. [70][71][72][73][74] Although both strategies appear to be attractive methods, they are characterized by some limitations including the incomplete representation of the whole embryonic genome, the potential maternal DNA contamination, the poor nucleic acid integrity and the unknown sampling time points to obtain acceptable amplification rates. 75 Well-designed studies are needed to evaluate the efficacy of cell-free DNA employment on clinical outcomes.…”

Section: Invasive and No Standardized Proceduresmentioning

confidence: 99%

Assessment of pre‐implantation genetic testing for embryo aneuploidies: A SWOT analysis

et al. 2019

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The recently re‐named pre‐implantation genetic testing for determining embryo aneuploidies (PGT‐A) is presently very popular although its acceptance by the scientific community is controversial. This approach still encounters drawbacks. This paper uses a SWOT (strengths, weaknesses, opportunities and threats) analysis to discuss salient points to be considered when examining the pre‐implantation genetic testing (PGT‐A) strategy to gather information from a range of perspectives. One of the strengths associated with the procedure is represented by an increase in implantation rate although data from the highest level of evidence do not support an increase in cumulative pregnancy rates. The current difficulty in the management of mosaicisms represents a weakness of PGT‐A. The application of the strategy represents an opportunity to favor the single embryo transfer while other advantages, such as reduction of time to pregnancy and emotional distress are controversial. Potential important threats, at present still undefined, are represented by the biopsy‐related damage to the blastocyst and the impact on neonatal and long‐term outcomes.

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Extracellular embryo genomic DNA and its potential for genotyping applications

Cited by 56 publications

References 19 publications

Noninvasive chromosome screening of human embryos by genome sequencing of embryo culture medium for in vitro fertilization

Noninvasive chromosome screening of human embryos by genome sequencing of embryo culture medium for in vitro fertilization

Pre-implantation genetic diagnosis and screening: now and the future

Assessment of pre‐implantation genetic testing for embryo aneuploidies: A SWOT analysis

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