Abstract
Preimplantation genetic screening (PGS) detects chromosomal aneuploidy from DNA extracted from trophectodermal biopsy of the embryos before implantation. Although a controlled study showed no difference in pregnancy rates between this invasive cell biopsy technique and a non-biopsied control group, the potential long-term damage by the current PGS method has not be completely ruled out. We therefore tested a less-invasive protocol which utilizes spent culture medium combining with blastocoel fluid (ECB) to assess chromosomal aneuploidy. We compared the new protocol with the currently employed trophectodermal biopsy method against chromosomal information obtained from the remaining embryo. We found that the new technique generated information about aneuploidy that was not entirely identical to obtained from the biopsied trophectoderm or the remaining embryo. As the origins of the DNA extracted from the three sample types were not the same, the significance and interpretation of each result would have its own meaning. The possible implications derived from the ECB results as well as those from cell biopsy were discussed. The effectiveness of this new approach in selecting the best embryo for uterine implantation awaits further long term evaluation.
Highlights
The controlled study by Scott et al.[15] showed no difference in pregnancy rates in comparison with a non-biopsied control group in humans, animal studies showed that embryo biopsy influenced epigenetic reprogramming during early embryo development impacting neural development and function in resulting mice[16]
An attempt of non-invasive Preimplantation genetic screening (PGS) was reported[23] in which the authors performed whole genome PCR amplification of DNA extracted from spent embryo culture medium and examined with NGS. They reported that the non-invasive technique obtained a high correlation to that obtained from the biopsy for detection of chromosomal aneuploidy but no comparison was made against the whole embryo or the remaining embryo
DNA concentrations in the embryo culture medium/blastocoel fluid (ECB) were sufficiently high for DNA amplification, NGS and aneuploidy analysis
Summary
The controlled study by Scott et al.[15] showed no difference in pregnancy rates in comparison with a non-biopsied control group in humans, animal studies showed that embryo biopsy influenced epigenetic reprogramming during early embryo development impacting neural development and function in resulting mice[16]. An attempt of non-invasive PGS was reported[23] in which the authors performed whole genome PCR amplification of DNA extracted from spent embryo culture medium and examined with NGS. They reported that the non-invasive technique obtained a high correlation to that obtained from the biopsy for detection of chromosomal aneuploidy (sensitivity: 88% and specificity: 84% respectively) but no comparison was made against the whole embryo or the remaining embryo. We report the development of a less-invasive technique using spent embryo culture medium/blastocoel fluid (ECB) to assess aneuploidy in the embryo and performed a comparative investigation of chromosomal analyses using ECB, biopsied cells and the remaining embryos. Following DNA extraction we employed a newly developed single cell DNA amplification protocol and NGS to evaluate the pros and cons of this new and less-invasive technique for possible embryo selection
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