Papers by Valeriy Kuznyetsov
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Reproductive BioMedicine Online, 2005
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Reproductive BioMedicine Online
Introduction: Some mosaic blastocysts are capable of forming viable normal pregnancies. However, ... more Introduction: Some mosaic blastocysts are capable of forming viable normal pregnancies. However, the association between mosaicism in trophectoderm (TE) and the inner cell mass (ICM) is far from being established. Unlike trophectoderm biopsy, non-invasive or minimally invasive preimplantation genetic testing for aneuploidy may better represent the entire blastocyst since cell-free embryonic DNA is most likely released into blastocoel fluid (BF) and spent embryo culture medium (SEM) from both TE and ICM.
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Reproductive Biomedicine Online , 2022
Introduction: Some mosaic blastocysts are capable of forming viable normal pregnancies. However, ... more Introduction: Some mosaic blastocysts are capable of forming viable normal pregnancies. However, the association between mosaicism in trophectoderm (TE) and the inner cell mass (ICM) is far from being established. Unlike trophectoderm biopsy, non-invasive or minimally invasive preimplantation genetic testing for aneuploidy may better represent the entire blastocyst since cell-free embryonic DNA is most likely released into blastocoel fluid (BF) and spent embryo culture medium (SEM) from both TE and ICM.
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Reproductive Biomedicine Online , 2022
Introduction: Some mosaic blastocysts are capable of forming viable normal pregnancies. However, ... more Introduction: Some mosaic blastocysts are capable of forming viable normal pregnancies. However, the association between mosaicism in trophectoderm (TE) and the inner cell mass (ICM) is far from being established. Unlike trophectoderm biopsy, non-invasive or minimally invasive preimplantation genetic testing for aneuploidy may better represent the entire blastocyst since cell-free embryonic DNA is most likely released into blastocoel fluid (BF) and spent embryo culture medium (SEM) from both TE and ICM.
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Systems biology in reproductive medicine, 2015
Despite broad utilization of sperm cryopreservation, little progress has been made to modify free... more Despite broad utilization of sperm cryopreservation, little progress has been made to modify freezing protocols or to improve rates of sperm survival. Vitrification is an alternative method for freezing human spermatozoa without toxic permeable cryoprotectants (CPAs). The purpose of our study was to optimize the vitrification and post thaw recovery of a small number of spermatozoa using only nonpermeating CPAs in a closed straw system in normozoospermic and severely oligozoospermic samples. Individual motile spermatozoa (n = 295) were selected from semen samples of 15 normozoospermic and 10 severe oligozoospermia patients. Overall sperm recovery after vitrification was 80% (n = 236) with 80% (n = 189) viability and 41.5% (n = 98) retained post-warming motility. Two different loading techniques were compared to transfer selected spermatozoa into straws in preparation for vitrification: by spontaneous capillary action (CA) and with the aid of a polar body biopsy (PBB) pipette. There w...
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CoGEN 2016 http://cogen2016.cme-congresses.com/posters.aspx List of Posters ABSTRACT no.10
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Scientific Reports
Preimplantation genetic testing for aneuploidies (PGT-A) using trophectoderm (TE) biopsy samples ... more Preimplantation genetic testing for aneuploidies (PGT-A) using trophectoderm (TE) biopsy samples is labour intensive, invasive, and subject to sampling bias. In this study, we report on the efficacy and factors affecting accuracy of a technique we pioneered for minimally invasive preimplantation genetic testing for aneuploidy (miPGT-A). Our technique uses cell-free embryonic DNA (cfeDNA) in spent embryo culture medium (SEM) combined with blastocoel fluid (BF) to increase the amount of assayable cfeDNA. We compared miPGT-A results (n = 145 embryos) with standard PGT-A analysis of the corresponding trophectoderm biopsy. We found that accuracy of miPGT was not related to blastocyst morphological grade. The overall concordance rate per sample for euploidy/aneuploidy status between miPGT-A and TE biopsy samples was 88/90 (97.8%), and was not different between good 47/48 (97.9%) and moderate/low quality blastocysts 41/42 (97.9%) (p > 0.05). Importantly, we also discovered that for cfeD...
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Reproductive BioMedicine Online
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Fertility and Sterility
OBJECTIVE: Blastocoel fluid (BF) and spent embryo culture medium (SEM) both contain cell-free emb... more OBJECTIVE: Blastocoel fluid (BF) and spent embryo culture medium (SEM) both contain cell-free embryonic DNA (cfeDNA). Attempts to use cfeDNA for non-invasive preimplantation genetic testing (NIPGT), brings to light several factors that could affect the accuracy of this approach. These include maternal contamination by cumulus/corona cells and DNA degradation. The objective of this study was to determine the accuracy, efficacy and reliability of whole genome amplification (WGA) to determine ploidy status (euploidy/aneuploidy) of the blastocyst using combined SEM+BF samples with or without using a cell lysis step before WGA.
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ABSTRACTImproved embryo selection is crucial in optimizing the results from assisted reproduction... more ABSTRACTImproved embryo selection is crucial in optimizing the results from assisted reproduction. Preimplantation genetic screening reduces time to pregnancy and miscarriages. Correlating the transcriptome of an embryo, with fertility treatments and outcomes, holds promise in improving the overall results. We developed a novel method for embryo selection in fertility treatments that integrates embryonic genomic and transcriptomic data and evaluated it in this pilot study.A total of 21 embryos donated for research were included. Three were used for the initial development and optimization of sample processing and sequencing. Thereafter, 18 embryos were used to demonstrate the clinical safety and reproducibility of our method. Two trophectoderm biopsies were taken from each embryo: one was processed as a clinical sample for genomic profiling (control, n=18), while the other biopsy (n=18) was split and utilized for independent, simultaneous genomic and transcriptomic analysis, here te...
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Scientific Reports
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Fertility and Sterility, 2016
Fertility and Sterility, September 2016, Volume 106, Issue 3, Supplement, Page e297 P-507
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Reproductive BioMedicine Online, 2003
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Reproductive BioMedicine Online, 2005
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PloS one, 2018
To assess whether embryonic DNA isolated from blastocyst culture conditioned medium (BCCM) combin... more To assess whether embryonic DNA isolated from blastocyst culture conditioned medium (BCCM) combined with blastocoel fluid (BF) could be used for blastocyst stage non-invasive preimplantation genetic testing for chromosomal aneuploidy (non-invasive preimplantation genetic screening, NIPGS). 47 embryos from 35 patients undergoing IVF. DNA analysis of combined BCCM plus BF in comparison with trophectoderm (TE) biopsy and/or whole blastocyst (WB)using next generation sequencing (NGS). Embryonic DNA was successfully amplified in 47/47 NIPGS samples (28 frozen-thawed and 19 fresh culture samples) ranging from 6.3 to 44.0 ng/μl. For frozen-thawed embryos, the concordance rate for whole chromosome copy number per sample was equivalent between NIPGS vs. TE biopsy, NIPGS vs. WB and TE vs. WB samples taken from the same embryo was 87.5%; 96.4% and 91.7% respectively (P>0.05), and the rate of concordance per single chromosome was 99.3%, 99.7% and 99.7%, respectively (P>0.05). In fresh cas...
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Fertility and Sterility
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Scientific Journal of Biology, 2020
The ability to select human embryo with the highest implantation potential remains one of the gre... more The ability to select human embryo with the highest implantation potential remains one of the greatest challenges in the management
of In Vitro Fertilization patients. Here, we review current methods for embryo selection including blastocyst static morphological grading,
analysis of embryo morphokinetic parameters, and preimplantation genetic testing for aneuploidy, and their impact on the success of
clinical outcomes. We also discuss emerging non-invasive cell-free embryonic DNA examination of spent embryo culture media. Other
experimental methods, such as metabolomics, proteomics, cumulus cells or trophectoderm biopsy sample transcriptomics remain
controversial and not yet suitable for routine clinical application. These are thus beyond the scope of this review. Traditionally, embryos with superior morphology have been selected for transfer, but these static parameters have been difficult to measure objectively and to use for prediction of blastocyst implantation potential. The introduction of time-lapse imaging system and morphokinetic markers to the laboratory practice allowed to study embryo cell dynamics during early preimplantation development, enhancing the evaluation of embryo quality. Nevertheless, morphokinetic annotations and predictive algorithms might vary between clinics because of different
culture environments, stimulation protocols and embryo developmental hallmarks, which overall limit their prognostic value for assessing
embryo viability. Moreover, the use of an objective but invasive technique of preimplantation genetic testing for embryo aneuploidy has
been shown to improve the selection of embryos with the most potential to implant and produce a healthy pregnancy. However, not all euploid embryos implant, likely owing to embryonic, endometrial factors, or both. Thus, at present, the combination of blastocyst testing for chromosomal abnormalities, analysis of embryo morphokinetics and morphological blastocyst scoring, as integrated approaches, will help to select developmentally competent embryos to maximize chances of successful reproductive outcome. Furthermore, implementing artificial intelligence to the field of Assisted Reproductive Technology may standardize the embryo selection process for a more reliable prediction of embryo quality and fate.
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Scientific Journal of Biology, 2020
The ability to select human embryo with the highest implantation potential remains one of the gre... more The ability to select human embryo with the highest implantation potential remains one of the greatest challenges in the management
of In Vitro Fertilization patients. Here, we review current methods for embryo selection including blastocyst static morphological grading,
analysis of embryo morphokinetic parameters, and preimplantation genetic testing for aneuploidy, and their impact on the success of
clinical outcomes. We also discuss emerging non-invasive cell-free embryonic DNA examination of spent embryo culture media. Other
experimental methods, such as metabolomics, proteomics, cumulus cells or trophectoderm biopsy sample transcriptomics remain
controversial and not yet suitable for routine clinical application. These are thus beyond the scope of this review. Traditionally, embryos with superior morphology have been selected for transfer, but these static parameters have been difficult to measure objectively and to use for prediction of blastocyst implantation potential. The introduction of time-lapse imaging system and morphokinetic markers to the laboratory practice allowed to study embryo cell dynamics during early preimplantation development, enhancing the evaluation
of embryo quality. Nevertheless, morphokinetic annotations and predictive algorithms might vary between clinics because of different
culture environments, stimulation protocols and embryo developmental hallmarks, which overall limit their prognostic value for assessing
embryo viability. Moreover, the use of an objective but invasive technique of preimplantation genetic testing for embryo aneuploidy has been shown to improve the selection of embryos with the most potential to implant and produce a healthy pregnancy. However, not all euploid embryos implant, likely owing to embryonic, endometrial factors, or both. Thus, at present, the combination of blastocyst testing for chromosomal abnormalities, analysis of embryo morphokinetics and morphological blastocyst scoring, as integrated approaches, will help to select developmentally competent embryos to maximize chances of successful reproductive outcome. Furthermore, implementing
artificial intelligence to the field of Assisted Reproductive Technology may standardize the embryo selection process for a more reliable
prediction of embryo quality and fate.
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Papers by Valeriy Kuznyetsov
of In Vitro Fertilization patients. Here, we review current methods for embryo selection including blastocyst static morphological grading,
analysis of embryo morphokinetic parameters, and preimplantation genetic testing for aneuploidy, and their impact on the success of
clinical outcomes. We also discuss emerging non-invasive cell-free embryonic DNA examination of spent embryo culture media. Other
experimental methods, such as metabolomics, proteomics, cumulus cells or trophectoderm biopsy sample transcriptomics remain
controversial and not yet suitable for routine clinical application. These are thus beyond the scope of this review. Traditionally, embryos with superior morphology have been selected for transfer, but these static parameters have been difficult to measure objectively and to use for prediction of blastocyst implantation potential. The introduction of time-lapse imaging system and morphokinetic markers to the laboratory practice allowed to study embryo cell dynamics during early preimplantation development, enhancing the evaluation of embryo quality. Nevertheless, morphokinetic annotations and predictive algorithms might vary between clinics because of different
culture environments, stimulation protocols and embryo developmental hallmarks, which overall limit their prognostic value for assessing
embryo viability. Moreover, the use of an objective but invasive technique of preimplantation genetic testing for embryo aneuploidy has
been shown to improve the selection of embryos with the most potential to implant and produce a healthy pregnancy. However, not all euploid embryos implant, likely owing to embryonic, endometrial factors, or both. Thus, at present, the combination of blastocyst testing for chromosomal abnormalities, analysis of embryo morphokinetics and morphological blastocyst scoring, as integrated approaches, will help to select developmentally competent embryos to maximize chances of successful reproductive outcome. Furthermore, implementing artificial intelligence to the field of Assisted Reproductive Technology may standardize the embryo selection process for a more reliable prediction of embryo quality and fate.
of In Vitro Fertilization patients. Here, we review current methods for embryo selection including blastocyst static morphological grading,
analysis of embryo morphokinetic parameters, and preimplantation genetic testing for aneuploidy, and their impact on the success of
clinical outcomes. We also discuss emerging non-invasive cell-free embryonic DNA examination of spent embryo culture media. Other
experimental methods, such as metabolomics, proteomics, cumulus cells or trophectoderm biopsy sample transcriptomics remain
controversial and not yet suitable for routine clinical application. These are thus beyond the scope of this review. Traditionally, embryos with superior morphology have been selected for transfer, but these static parameters have been difficult to measure objectively and to use for prediction of blastocyst implantation potential. The introduction of time-lapse imaging system and morphokinetic markers to the laboratory practice allowed to study embryo cell dynamics during early preimplantation development, enhancing the evaluation
of embryo quality. Nevertheless, morphokinetic annotations and predictive algorithms might vary between clinics because of different
culture environments, stimulation protocols and embryo developmental hallmarks, which overall limit their prognostic value for assessing
embryo viability. Moreover, the use of an objective but invasive technique of preimplantation genetic testing for embryo aneuploidy has been shown to improve the selection of embryos with the most potential to implant and produce a healthy pregnancy. However, not all euploid embryos implant, likely owing to embryonic, endometrial factors, or both. Thus, at present, the combination of blastocyst testing for chromosomal abnormalities, analysis of embryo morphokinetics and morphological blastocyst scoring, as integrated approaches, will help to select developmentally competent embryos to maximize chances of successful reproductive outcome. Furthermore, implementing
artificial intelligence to the field of Assisted Reproductive Technology may standardize the embryo selection process for a more reliable
prediction of embryo quality and fate.
of In Vitro Fertilization patients. Here, we review current methods for embryo selection including blastocyst static morphological grading,
analysis of embryo morphokinetic parameters, and preimplantation genetic testing for aneuploidy, and their impact on the success of
clinical outcomes. We also discuss emerging non-invasive cell-free embryonic DNA examination of spent embryo culture media. Other
experimental methods, such as metabolomics, proteomics, cumulus cells or trophectoderm biopsy sample transcriptomics remain
controversial and not yet suitable for routine clinical application. These are thus beyond the scope of this review. Traditionally, embryos with superior morphology have been selected for transfer, but these static parameters have been difficult to measure objectively and to use for prediction of blastocyst implantation potential. The introduction of time-lapse imaging system and morphokinetic markers to the laboratory practice allowed to study embryo cell dynamics during early preimplantation development, enhancing the evaluation of embryo quality. Nevertheless, morphokinetic annotations and predictive algorithms might vary between clinics because of different
culture environments, stimulation protocols and embryo developmental hallmarks, which overall limit their prognostic value for assessing
embryo viability. Moreover, the use of an objective but invasive technique of preimplantation genetic testing for embryo aneuploidy has
been shown to improve the selection of embryos with the most potential to implant and produce a healthy pregnancy. However, not all euploid embryos implant, likely owing to embryonic, endometrial factors, or both. Thus, at present, the combination of blastocyst testing for chromosomal abnormalities, analysis of embryo morphokinetics and morphological blastocyst scoring, as integrated approaches, will help to select developmentally competent embryos to maximize chances of successful reproductive outcome. Furthermore, implementing artificial intelligence to the field of Assisted Reproductive Technology may standardize the embryo selection process for a more reliable prediction of embryo quality and fate.
of In Vitro Fertilization patients. Here, we review current methods for embryo selection including blastocyst static morphological grading,
analysis of embryo morphokinetic parameters, and preimplantation genetic testing for aneuploidy, and their impact on the success of
clinical outcomes. We also discuss emerging non-invasive cell-free embryonic DNA examination of spent embryo culture media. Other
experimental methods, such as metabolomics, proteomics, cumulus cells or trophectoderm biopsy sample transcriptomics remain
controversial and not yet suitable for routine clinical application. These are thus beyond the scope of this review. Traditionally, embryos with superior morphology have been selected for transfer, but these static parameters have been difficult to measure objectively and to use for prediction of blastocyst implantation potential. The introduction of time-lapse imaging system and morphokinetic markers to the laboratory practice allowed to study embryo cell dynamics during early preimplantation development, enhancing the evaluation
of embryo quality. Nevertheless, morphokinetic annotations and predictive algorithms might vary between clinics because of different
culture environments, stimulation protocols and embryo developmental hallmarks, which overall limit their prognostic value for assessing
embryo viability. Moreover, the use of an objective but invasive technique of preimplantation genetic testing for embryo aneuploidy has been shown to improve the selection of embryos with the most potential to implant and produce a healthy pregnancy. However, not all euploid embryos implant, likely owing to embryonic, endometrial factors, or both. Thus, at present, the combination of blastocyst testing for chromosomal abnormalities, analysis of embryo morphokinetics and morphological blastocyst scoring, as integrated approaches, will help to select developmentally competent embryos to maximize chances of successful reproductive outcome. Furthermore, implementing
artificial intelligence to the field of Assisted Reproductive Technology may standardize the embryo selection process for a more reliable
prediction of embryo quality and fate.
niPGT-A) techniques. The objectives of this study were to evaluate if determining the ploidy status of non-invasive SBM+BF samples would impact the pregnancy rates after the eSFET of blastocysts designated as euploid by TE sampling.
viable normal pregnancies. However, the association between mosaicism
in trophectoderm (TE) and the inner cell mass (ICM) is far from being
established. Unlike trophectoderm biopsy, non-invasive or minimally
invasive preimplantation genetic testing for aneuploidy (miPGT-A) may better represent the entire blastocyst since cell-free embryonic DNA is most likely released into blastocoel fluid (BF) and spent embryo culture medium (SEM) from both TE and ICM.
There is growing interest in the use of minimally invasive and non-invasive preimplantation genetic testing (mi/NIPGT) based on cell-free embryonic DNA (cfeDNA) from blastocoel fluid (BF) or/and spent embryo culture medium (SEM). However, despite the promising results, concordance rate for ploidy between cfeDNA and trophectoderm (TE) biopsy, inner cell mass (ICM) and/or whole blastocysts is still controversial issue. Moreover, it is still debated origin of cell-free embryonic DNA in spent medium from
human embryo (TE DNA or both TE DNA and ICM DNA). The objective of this study was to evaluate the efficacy and accuracy of NIPGT-A using cfeDNA in combined SEM+BF samples in comparison to corresponding TE biopsy PGT-A.
There is growing interest in the use of minimally invasive and non-invasive preimplantation genetic testing (mi/NIPGT) based on cell-free embryonic DNA (cfeDNA) from blastocoel fluid (BF) or/and spent embryo culture medium (SEM). However, despite the promising results, concordance rate for ploidy between cfeDNA and trophectoderm (TE) biopsy, inner cell mass (ICM) and/or whole blastocysts is still controversial issue. Moreover, it is still debated origin of cell-free embryonic DNA in spent medium from
human embryo (TE DNA or both TE DNA and ICM DNA). The objective of this study was to evaluate the efficacy and accuracy of NIPGT-A using cfeDNA in combined SEM+BF samples in comparison to corresponding TE biopsy PGT-A.
permeable cryoprotectants might be an attractive alternative to the vapor freezing for testicular or epididymal spermatozoa, or severe oligozoospermic samples. The objective of this study was to evaluate the testicular and epididymal sperm motility and viability after using our novel technique of permeable cryoprotectant-free sperm vitrification and to analyze the embryo development after ICSI with these vitrified spermatozoa.
There is growing interest in the use of minimally invasive and non-invasive preimplantation genetic testing (mi/NIPGT) based on cell-free embryonic DNA (cfeDNA) from blastocoel fluid (BF) or/and spent embryo culture medium (SEM). However, despite the promising results, concordance rate for ploidy between cfeDNA and trophectoderm (TE) biopsy, inner cell mass (ICM) and/or whole blastocysts is still controversial issue. Moreover, it is still debated origin of cell-free embryonic DNA in spent medium from
human embryo (TE DNA or both TE DNA and ICM DNA). The objective of this study was to evaluate the efficacy and accuracy of NIPGT-A using cfeDNA in combined SEM+BF samples in comparison to corresponding TE biopsy PGT-A.
There is growing interest in the use of minimally invasive and non-invasive preimplantation genetic testing (mi/NIPGT) based on cell-free embryonic DNA (cfeDNA) from blastocoel fluid (BF) or/and spent embryo culture medium (SEM). However, despite the promising results, concordance rate for ploidy between cfeDNA and trophectoderm (TE) biopsy, inner cell mass (ICM) and/or whole blastocysts is still controversial issue. Moreover, it is still debated origin of cell-free embryonic DNA in spent medium from human embryo (TE DNA or both TE DNA and ICM DNA). The objective of this study was to evaluate the efficacy and accuracy of NIPGT-A using cfeDNA in combined SEM+BF samples in comparison to corresponding TE biopsy PGT-A.
- Blastocyst biopsy is labour intensive and require highly trained embryologists
- TE biopsy is invasive and may have a negative impact on implantation and clinical pregnancy
- TE biopsy may not accurately represent the inner cell mass (ICM) and the reminder of trophectoderm (TE)
- Long-term sequelae of TE biopsy are presently unknown (Zhang et al., 2019; Tocci et al., 2020; Makhijani et al., 2021)
(SEM) both contain cell-free embryonic DNA (cfeDNA). Attempts to
use cfeDNA for non-invasive preimplantation genetic testing (NIPGT),
brings to light several factors that could affect the accuracy of this
approach. These include maternal contamination by cumulus/corona cells and DNA degradation. The objective of this study was to determine the accuracy, efficacy and reliability of whole genome amplification
(WGA) to determine ploidy status (euploidy/aneuploidy) of the blastocyst using combined SEM+BF samples with or without using a cell lysis step before WGA.
(POC) can elucidate the reason for miscarriage in approximately 50-70%
of first trimester miscarriages. Assessment of the fetal chromosomal composition may be very helpful in counselling and management of patients experiencing miscarriages, especially after IVF, or in patients with recurrent pregnancy losses. However, obtaining fetal tissue from early miscarriages is often compromised by maternal cell contamination (MCC). Here we present the results from assessing early POC samples (<10 GW) after IVF single embryo transfer, controlling for MCC, using whole genome NGS at the CReATe Fertility Centre.
In addition, without permeable cryoprotectants, a washing step is not required, and warmed sperms can be placed directly into an ICSI dish.