Is Day-4 morula biopsy a feasible alternative for preimplantation genetic testing?

Orvieto, Raoul; Feldman, Baruch; Wiesel, Marine; Shani, Hagit; Aizer, Adva

doi:10.1371/journal.pone.0238599

Cited by 6 publications

(7 citation statements)

References 12 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In human in vitro embryo production, preimplantation genetic testing (PGT) is a common practice, especially for patients with a high risk of a genetic diseases [ 29 , 30 , 31 ]. For example, embryos might be screened for aneuploidy, which is the presence of an abnormal number of chromosomes in a cell, by analyzing biopsies of day-3 or day-4 morulae.…”

Section: Discussionmentioning

confidence: 99%

“…For example, embryos might be screened for aneuploidy, which is the presence of an abnormal number of chromosomes in a cell, by analyzing biopsies of day-3 or day-4 morulae. This allows the selection of a chromosomally normal embryo for transfer for the development of healthy offspring [ 29 , 30 , 31 ]. Therefore, good and minimally invasive molecular preimplantation testing methods and practices are essential in assisted reproduction.…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Detecting Embryo Developmental Potential by Single Blastomere RNA-Seq

Nõmm

Ivask

Pärn

et al. 2023

Genes

View full text Add to dashboard Cite

Recent advances in preimplantation embryo diagnostics enable a wide range of applications using single cell biopsy and molecular-based selection techniques without compromising embryo production. This study was conducted to develop a single cell embryo biopsy technique and gene expression analysis method with a very low input volume to ensure normal embryo development and to see if there are differences in gene expression profiles between day-5 biopsied bovine embryos that developed into blastocysts and embryos arrested at morula stage. Out of the 65 biopsied morulae, 32 developed to blastocysts (49.2%). Out of the 13580 successfully annotated genes, 1204 showed a difference in mRNA expression level. Out of these, 155 genes were expressed in embryos developing to blastocysts. The pathway enrichment analysis revealed significant enrichment in “organelle biogenesis and maintenance”, “mRNA splicing” and “mitochondrial translation” pathways. These findings suggest principal differences in gene expression patterns and functional networks of embryos able to reach the blastocyst stage compared to embryos arrested in development. Our preliminary data suggest that single blastomere biopsy and selected gene expression profiles at morula stage could offer additional possibilities for early preimplantation embryo selection before transfer.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Detecting Embryo Developmental Potential by Single Blastomere RNA-Seq

Nõmm

Ivask

Pärn

et al. 2023

Genes

View full text Add to dashboard Cite

show abstract

“…opening of the zona pellucida on day 3 of in vitro development, enabling the TE cells herniation through that opening and their biopsy on day 5. We might therefore suggest deferring the zona pellucida opening to Day-4 [ 11 ], enabling the retrieval of the cell debris/fragments for the molecular diagnosis. If the results of latter molecular diagnosis, on the following day (Day-5), will be complete/conclusive, the embryo is diagnosed and can be handled accordingly.…”

Section: Discussionmentioning

confidence: 99%

“…If the results of latter molecular diagnosis, on the following day (Day-5), will be complete/conclusive, the embryo is diagnosed and can be handled accordingly. Of notice, while comparing the efficacy and clinical outcome of PGT-M undertaken on Day-3 or Day-4 embryos, Day-4 embryo biopsy was found to be feasible and yielded comparable and even higher ongoing pregnancy rate if undertaken at the morula stage [ 11 ].…”

Section: Discussionmentioning

confidence: 99%

Can expelled cells/debris from a developing embryo be used for PGT?

et al. 2021

Self Cite

View full text Add to dashboard Cite

Background Preimplantation genetic testing (PGT) is offered to a wide range of structural and numerical chromosomal imbalances, with PGT- polymerase chain reaction (PCR), as the method of choice for amplifying the small DNA content achieved from the blastomere biopsy or trophectoderm (TE) biopsy, that might have a detrimental impact on embryonic implantation potential. Since human embryos cultured until Day-5–6 were noticed to expel cell debris/ fragments within the zona pellucida, we aimed to examine whether these cell debris/ fragments might be used for PGT, as an alternative to embryo biopsy. Methods Blastocysts, which their Day-3 blastomere biopsy revealed an affected embryo with single-gene defect, and following hatching leaved cell debris/fragments within the zona pellucida were analyzed. Each blastocyst and its corresponding cell debris/fragments were separated and underwent the same molecular analysis, based on multiplex PCR programs designed for haplotyping using informative microsatellites markers. The main outcome measure was the intra-embryo congruity of Day-3 blastomere biopsy and its corresponding blastocyst and cell debris/fragments. Results Fourteen affected embryos from 9 women were included. Only 8/14 (57.2%) of embryos demonstrated congruent molecular genetic results between Day-3 embryo and its corresponding blastocyst and cell debris/fragments. In additional 6/14 (42.8%) embryos, molecular results of the Day-3 embryos and their corresponding blastocysts were congruent, while the cell debris/fragments yielded no molecular diagnoses (incomplete diagnoses). Conclusions It might be therefore concluded, that in PGT cycles, examining the cell debris/fragments on Day-4, instead of Day-3 blastomere or Day-5 TE biopsies, is feasible and might avoid embryo biopsy with its consequent detrimental effect on embryos’ implantation potential. Whenever the latter results in incomplete diagnosis, TE biopsy should be carried out on Day-5 for final genetic results. Further large well-designed studies are required to validate the aforementioned PGT platform.

show abstract

“…Biopsy of day 4 embryos (morula), prior to the blastocyst stage, has also been described and permits the biopsy of more than one cell leaving time for genetic analysis and fresh embryo transfer [ 25 ]. Reports on this approach are limited and there remains uncertainty regarding the fate of cells retrieved and how biopsy at this embryonic stage may affect development and implantation [ 26 ]. Furthermore, the reproducibility and safety of this technique have not been systematically investigated and the procedure has seen very limited application to date [ 21 ].…”

Section: Traditional and Modern Sources Of Embryonic Dnamentioning

confidence: 99%

An Update on Non-invasive Approaches for Genetic Testing of the Preimplantation Embryo

Kakourou

Mamasa

Vrettou

et al. 2022

View full text Add to dashboard Cite

Preimplantation Genetic Testing (PGT) aims to reduce the chance of an affected pregnancy or improve success in an assisted reproduction cycle. Since the first established pregnancies in 1990, methodological approaches have greatly evolved, combined with significant advances in the embryological laboratory. The application of preimplantation testing has expanded, while accuracy and reliability of monogenic and chromosomal analysis has improved. The procedure traditionally employs an invasive approach to assess the nucleic acid content of embryos. All biopsy procedures require high technical skill, costly equipment, and may impact both the accuracy of genetic testing and embryo viability. To overcome these limitations, many researchers have focused on the analysis of cell-free DNA (cfDNA) at the preimplantation stage, sampled either from the blastocoel or from embryo culture media, to determine the genetic status of the embryo non-invasively. Studies have assessed the origin of cfDNA and application in non-invasive testing for monogenic disease and chromosomal aneuploidies. Herein, we discuss the state-of-the-art for modern non-invasive embryonic genetic material assessment, in the context of PGT. The results are difficult to integrate due to numerous methodological differences between the studies, while further work is required to assess the suitability of cfDNA analysis for clinical application.

show abstract

Is Day-4 morula biopsy a feasible alternative for preimplantation genetic testing?

Cited by 6 publications

References 12 publications

Detecting Embryo Developmental Potential by Single Blastomere RNA-Seq

Detecting Embryo Developmental Potential by Single Blastomere RNA-Seq

Can expelled cells/debris from a developing embryo be used for PGT?

An Update on Non-invasive Approaches for Genetic Testing of the Preimplantation Embryo

Contact Info

Product

Resources

About