Deoxyribonucleic acid detection in blastocoelic fluid: a new predictor of embryo ploidy and viable pregnancy

The presence of DNA in blastocoelic fluids negatively correlates with the blastocyst ploidy condition defined by trophectoderm cell biopsy and with the implantation potential of trophectoderm-euploid blastocysts.

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Volume 111, Issue 1, Pages 77–85

Authors:

M. Cristina Magli, M.Sc., Cristina Albanese, M.Sc., Andor Crippa, Ph.D., Carla Tabanelli, M.D., Anna P. Ferraretti, M.D., Luca Gianaroli, M.D.

Abstract:

Objective

To investigate blastocysts, defined as euploid and aneuploid by trophectoderm (TE) cell analysis, for the presence of DNA in the blastocoelic fluid (BF) detected by whole-genomic amplification (WGA); and to correlate the presence of DNA in BF with the clinical outcome after the transfer of TE-euploid blastocysts.

Design

Retrospective study.

Setting

In vitro fertilization unit.

Patient(s)

This study included 91 patients performing preimplantation genetic testing for aneuploidy on TE cells from January 2015 to December 2017. In the case of ET, only single blastocyst transfers were performed.

Intervention(s)

Blastocoelic fluids and TE cells were retrieved from 256 blastocysts before vitrification. All blastocysts were diagnosed by array-comparative genomic hybridization (a-CGH) on TE cells. Amplification and a-CGH of DNA from BFs was performed at a later time after TE biopsy and ET.

Main Outcome Measure(s)

Whole-genomic amplification of BFs, evaluation of the chromosome condition in BFs and TE cells, and correlation of BF results with the clinical outcome of TE-euploid transferred blastocysts.

Result(s)

The incidence of amplification after WGA was significantly lower in BFs from TE-euploid blastocysts (n = 32, 45%) when compared with the aneuploid ones (n = 150, 81%), resulting in 182 BFs with successful DNA amplification. When submitted to a-CGH, informative results were obtained from 172 BFs. Comparison of these results with those from the corresponding TE cells gave a ploidy concordance of 93.6% and a mean number of aneuploid events per sample that was higher in BFs than in TE cells (2.0 vs. 1.4, respectively). After the transfer of 53 TE-euploid blastocysts, the clinical pregnancy rate was 77% in the group with BF-failed amplification, and 37% after BF-successful amplification. The same trend was found for the ongoing pregnancy rate (68% vs. 31.5%, respectively).

Conclusion(s)

The presence of DNA in BFs detected by WGA is correlated with the blastocyst ploidy condition defined by TE cell biopsy and with the implantation potential of TE-euploid blastocysts. These findings could have a clinical implication for the selection of the most viable embryo for transfer because, after submitting BFs to WGA, priority would be given to TE-euploid blastocysts with BF-failed amplification. Similarly, BF-failed amplification could be an additional selection criterion to prioritize embryos for transfer even in conventional IVF cycles with blastocysts that were vitrified after BF aspiration.


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Fertility and Sterility® is an international journal for obstetricians, gynecologists, reproductive endocrinologists, urologists, basic scientists and others who treat and investigate problems of infertility and human reproductive disorders. The journal publishes juried original scientific articles in clinical and laboratory research relevant to reproductive endocrinology, urology, andrology, physiology, immunology, genetics, contraception, and menopause. Fertility and Sterility® encourages and supports meaningful basic and clinical research, and facilitates and promotes excellence in professional education, in the field of reproductive medicine.

2 Comments

Go to the profile of Andre Hazout
Andre Hazout about 1 month ago

I have red with interest the Paper of the Gianarolli team. I don't contest the results but the value of the free DNA assay into the blastocele and the results of the TE biopsy can not be compared. Even if there is a coincidence between the viability of blastocysts viability and the absence of DNA in their blastocele and  euploid blastocyst viability. Moreover Few centers can practice both explorations despite high cost


Go to the profile of Luca Gianaroli
Luca Gianaroli 7 days ago

There is a difference between coincidence and significant statistical
analysis. High costs are part of medical research. As this procedure is part
of a study and because of results obtained, costs will become much lower due
to the fact that a simple DNA amplification can suggest which embryo to
transfer first.