Loading…
Genomic DNA in human blastocoele fluid
Abstract IVF often requires embryo cryopreservation through vitrification. During the vitrification process, the embryos can be collapsed by withdrawing the blastocoele fluid. The metabolomic profile of blastocoele fluid has been recently investigated by high-performance liquid chromatography-electr...
Saved in:
Published in: | Reproductive biomedicine online 2013-06, Vol.26 (6), p.603-610 |
---|---|
Main Authors: | , , , , , , |
Format: | Article |
Language: | English |
Subjects: | |
Citations: | Items that this one cites Items that cite this one |
Online Access: | Get full text |
Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
Summary: | Abstract IVF often requires embryo cryopreservation through vitrification. During the vitrification process, the embryos can be collapsed by withdrawing the blastocoele fluid. The metabolomic profile of blastocoele fluid has been recently investigated by high-performance liquid chromatography-electrospray ionization-mass spectrometry to provide metabolite information that can help estimations of implantation efficiency. However, the presence of embryo DNA in blastocoele fluid has not been reported to date. This study shows using real-time PCR that genomic DNA was present in about 90% of blastocoele fluid samples harvested during the vitrification procedure. Moreover, the potential for determining embryo sex directly from blastocoele fluid is demonstrated by amplifying the multicopy genes TSPY1 (on the Y chromosome) and TBC1D3 (on chromosome 17). This opens up the possibility of screening embryos from couples carrying an X-linked disorder to identify male embryos at high risk of disease. The application of whole-genome amplification technologies to fluid samples is also shown to be feasible, potentially allowing more comprehensive genetic tests. As proof of principle, microarray comparative genomic hybridization was attempted to confirm the sex of embryos as well as detect several aneuploidies. However, further studies are needed to validate this approach and confirm that the accuracy is sufficient for diagnostic purposes. IVF offers personal approaches to couples according to their specific infertility problems. IVF includes a preimplantation genetic diagnosis, single-embryo transfer, blastocyst transfer and optimization of uterine receptivity. These procedures require a valid embryo cryopreservation system. Vitrification is a cryopreservation method that offers superior survival and pregnancy rates performing an artificial shrinkage of a fully expanded blastocyst. This procedure can be performed with different methods, including laser-pulse, repeated micropipetting, microneedle puncture or microsuction, and there is a significant improvement in survival rates when blastocysts were collapsed. With microsuction, the blastocoele fluid is withdrawn and (typically) discarded. The aim of this study was to investigate the presence of DNA in blastocoele fluid. Recently, the metabolomic profile of blastocoele fluid was characterized, but to date, no studies have reported evidence of DNA. Here, using a real-time PCR-based approach, we showed that genomic DNA was am |
---|---|
ISSN: | 1472-6483 1472-6491 |
DOI: | 10.1016/j.rbmo.2013.02.012 |