Non-invasive preimplantation genetic screening using array comparative genomic hybridization on spent culture media: a proof-of-concept pilot study

Feichtinger, Michael; Vaccari, Enrico; Carli, Luca; Wallner, Elisabeth I.; Mädel, Ulrike; Figl, Katharina; Palini, Simone; Feichtinger, Wilfried

doi:10.1016/j.rbmo.2017.03.015

Cited by 75 publications

(80 citation statements)

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“…Nonetheless, blastocentesis entails some degree of invasiveness to the embryo, although it is minimal. To develop a completely noninvasive, inexpensive, and easily implementable methodology for PGT purposes, several investigators proposed the use of spent culture medium collected at blastocyst stage (SBM) as a source of embryonic DNA (15)(16)(17). In particular, Xu et al (18) have shown high ploidy concordance rate between SBM samples and their corresponding embryos (>80%).…”

mentioning

confidence: 99%

Diagnostic efficacy of blastocoel fluid and spent media as sources of DNA for preimplantation genetic testing in standard clinical conditions

Capalbo

Romanelli

Patassini

et al. 2018

Fertility and Sterility

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Objective: To determine whether blastocoel fluid (BF) or spent blastocyst medium (SBM) is a suitable template for genotype and/or karyotype assessment of in vitro fertilization-generated embryos. Design: Prospective blinded study. Setting: Genetic laboratory. Patient(s): From 26 patients undergoing preimplantation genetic testing (PGT) treatments, 103 trophectoderms (TE), 92 BF samples, and 72 SBM samples. Intervention(s): The BF and SBM were retrieved at the time of TE biopsy. Two DNA extraction strategies were evaluated on independent BF and SBM samples. Further enrolled samples were processed using next-generation sequencing and quantitative polymerase chain reaction for assessment of monogenic disorders (PGT-M) or aneuploidy (PGT-A). Main Outcome Measure(s): DNA amplification and concordance rates across BF, SBM, and TE to assess diagnostic efficiency. Result(s): No differences were detected among the DNA extraction methods tested. In PGT-M tests, for BF and SBM, 2.9% and 20.8% of all samples, respectively, produced a diagnosis concordant with the corresponding TE (n ¼ 2 of 69 and 15 of 72, respectively). The SBM samples were associated with higher discordance rates and higher artifacts/contamination detection compared with BF. In multiple occasions, the maternal mutated variant was detected in the SBM of homozygous wild-type embryos, showing evidence of maternal DNA persistence in culture medium. In PGT-A tests, BF analysis showed high amplification failure rates (65.2%) and an overall concordance rate of 37.5% among amplified samples. Conclusion(s): Based on current methodologies, BF and SBM genetic analyses do not provide sufficiently reliable results to be employed clinically. Until the risk of maternal contamination can be properly prevented, SBM should not be used for PGT-M purposes. (Fertil Steril Ò 2018;110:870-9. Ó2018 by American Society for Reproductive Medicine.) El resumen está disponible en Español al final del artículo.

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mentioning

confidence: 99%

Diagnostic efficacy of blastocoel fluid and spent media as sources of DNA for preimplantation genetic testing in standard clinical conditions

Capalbo

Romanelli

Patassini

et al. 2018

Fertility and Sterility

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“…The sample size ranged from seven (Shamonki et al, 2016;Liu et al, 2017) to 70 (Hammond et al, 2017) couples undergoing assisted reproductive technology procedures. Gamete fertilization was carried out mainly by intracytoplasmic sperm injection (ICSI) (86.7% [13 out of 15]) (Stigliani et al, 2013;Wu et al, 2015;Shamonki et al, 2016;Xu et al, 2016;Feichtinger et al, 2017;Hammond et al, 2017;Liu et al, 2017;Yang et al, 2017;Capalbo et al, 2018;Ho et al, 2018;Kuznyetsov et al, 2018;Li et al, 2018;Vera-Rodriguez et al, 2018) and the other two studies did not report the fertilization method Galluzzi et al, 2015). In 86.6% (13 out of 15) of studies, Cf-DNA from SCM alone was evaluated (Stigliani et al, 2013;Assou et al, 2014;Galluzzi et al, 2015;Wu et al, 2015;Shamonki et al, 2016;Xu et al, 2016;Feichtinger et al, 2017;Liu et al, 2017;Capalbo et al, 2018;Ho et al, 2018;Vera-Rodriguez et al, 2018), whereas a mixture of SCM and blastocoel fluid was assessed in 26.7% (four out of 15) of them (Hammond et al, 2017;Yang et al, 2017;Kuznyetsov et al, 2018;Li et al, 2018).…”

Section: Study Characteristicsmentioning

confidence: 99%

“…The characteristics of the embryos associated with each SCM sample were also heterogeneous. Specifically, 93.3% (14 out of 15) studies evaluated cf-DNA released from fresh embryos (Stigliani et al, 2013;Galluzzi et al, 2015;Wu et al, 2015;Shamonki et al, 2016;Xu et al, 2016;Hammond et al, 2017;Liu et al, 2017;Yang et al, 2017;Feichtinger et al, 2017;Capalbo et al, 2018;Ho et al, 2018;Kuznyetsov et al, 2018;Li et al, 2018;Vera-Rodriguez et al, 2018), and 20.0% (three out of 15) of studies Kuznyetsov et al, 2018;Ho et al, 2018) assessed cf-DNA secreted from cryopreserved embryos. Moreover, embryo micromanipulation, e.g.…”

Section: Study Characteristicsmentioning

confidence: 99%

“…Moreover, embryo micromanipulation, e.g. assisted hatching and blastocoel collapse or embryo biopsy, was carried out before SCM sampling in 60.0% (nine out of 15) of studies Shamonki et al, 2016;Feichtinger et al, 2017;Hammond et al, 2017;Yang et al, 2017;Ho et al, 2018;Kuznyetsov et al, 2018;Li et al, 2018; Vera-Rodriguez FIGURE 2 Flow chart of search strategy and study inclusion (Moher et al, ). et al, 2018.…”

Section: Study Characteristicsmentioning

confidence: 99%

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Is cell-free DNA in spent embryo culture medium an alternative to embryo biopsy for preimplantation genetic testing? A systematic review

Brouillet

Martinez

Coutton

et al. 2020

Reproductive BioMedicine Online

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Spent culture medium (SCM) represents a possible source of embryonic DNA for non-invasive preimplantation genetic testing (PGT). Before considering routine clinical use, methodologies need to be improved to increase detection of embryo DNA in SCM while preventing contamination by exogenous and maternal DNA. The identification of DNA origin (in particular, maternal versus embryonic) is necessary for improved reliability of SCM-PGT. ABSTRACTPreimplantation genetic testing (PGT) is increasingly used worldwide. It currently entails the use of invasive techniques, i.e. polar body, blastomere, trophectoderm biopsy or blastocentesis, to obtain embryonic DNA, with major technical limitations and ethical issues. Evidence suggests that invasive PGT can lead to genetic misdiagnosis in the case of embryo mosaicism, and, consequently, to the selection of affected embryos for implantation or to the destruction of healthy embryos. Recently, spent culture medium (SCM) has been proposed as an alternative source of embryonic DNA. An increasing number of studies have reported the detection of cell-free DNA in SCM and highlighted the diagnostic potential of non-invasive SCM-based PGT for assessing the genetic status of preimplantation human embryos obtained by IVF. The reliability of this approach for clinical applications, however, needs to be determined. In this systematic review, published evidence on non-invasive SCM-based PGT is presented, and its current benefits and limitations compared with invasive PGT. Then, ways of optimizing and standardizing procedures for non-invasive SCM-based PGT to prevent technical biases and to improve performance in future studies are discussed. Finally, clinical perspectives of non-invasive PGT are presented and its future applications in reproductive medicine highlighted.

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“…Whether the blastocoele fluid truly represents the chromosome complement of the whole embryo still needs to be demonstrated; however, this is an interesting research avenue as blastocysts completely recover and regain their original morphology after biopsy, and blastocoele aspiration is therefore considered to be less invasive than blastocyst biopsy [32]. Analysis of the culture medium in which the embryo was cultured is non-invasive, although here there are reasonable doubts whether the DNA present in the medium is coming from the embryo [33][34][35].…”

Section: Blastocyst Biopsymentioning

confidence: 99%

Preimplantation Genetic Screening

Sermon¹

2017

obm genet

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The main aim of PGS has always been to improve IVF outcome, especially in patient groups assumed to have higher rates of chromosomally abnormal embryos, such as patients of advanced maternal age. In that sense, PGS is quite different from other types of screening as discussed in other papers in this issue. Today it bears no doubt that blastocysts found to be uniformly aneuploid in a biopsy will fail to implant, or worse, will implant and lead to a pregnancy and birth carrying a major chromosomal abnormality, such as trisomy 21. However, it has been argued that a cohort of embryos cannot be improved, and that PGS is only a selection method for which efficiency has not been proven. PGS would never increase the live birth rate for that given cohort, even with a 100% efficiency rate of embryo cryopreservation. The current debate on whether PGS should be applied and to which patients it should be offered has shifted from the effect on live birth rates towards other outcomes such as the reduction of transfers and of miscarriages. Taking the undeniable higher cost of IVF into account when PGS is included, what is the benefit to the patient? The views on this question differ on whether PGS is an additional source of income for the IVF clinic and may or may not balance the extra cost for cryopreservation and embryo transfer for the patients, or whether society pays for IVF treatments and may decide not to want to invest in a medical act that does not improve the primary goal of IVF, i.e. having a healthy child. PGS is also often presented as diminishing patient anxiety and stress through decreasing unnecessary embryos transfers and miscarriages, although no data on this

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Non-invasive preimplantation genetic screening using array comparative genomic hybridization on spent culture media: a proof-of-concept pilot study

Cited by 75 publications

References 40 publications

Diagnostic efficacy of blastocoel fluid and spent media as sources of DNA for preimplantation genetic testing in standard clinical conditions

Diagnostic efficacy of blastocoel fluid and spent media as sources of DNA for preimplantation genetic testing in standard clinical conditions

Is cell-free DNA in spent embryo culture medium an alternative to embryo biopsy for preimplantation genetic testing? A systematic review

Preimplantation Genetic Screening

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