P–159 Slow-growing embryos should be frozen on day 5
Authors: Zamora M J , Katsouni I , Garcia D , Vassena R , Rodríguez A 
Published: August 2021

Abstract

Study question
What is the live birth rate after frozen embryo transfer (FET) of slow-growing embryos frozen on day 5 (D5) or on day 6 (D6)?

Summary answer
The live birth rate after single FET is significantly higher for slow-growing embryos frozen on D5 compared to those frozen on D6.

What is known already
Most data on the outcomes of blastocyst transfer stem from studies that evaluate fresh transfer from normal growing D5 blastocyst ET. However not all embryos will begin blastulation nor reach the fully expanded stage by D5; those are the slow-growing embryos. Studies that compare D5 to D6 embryos in FET cycles show contradictory results. Some have reported higher clinical pregnancy rates after D5 FET, while others have reported similar outcomes for D5 and D6 cryopreserved blastocyst transfers. There is a lack of evidence regarding the best approach for vitrifying embryos that exhibit a slow developmental kinetic.

Study design, size, duration
This retrospective cohort study included 821 single FET of slow-growing embryos frozen on D5 or D6, belonging to patients undergoing in vitro fertilization with donor oocytes between January 2011 and October 2019, in a single fertility center. The origin of blastocysts was either supernumerary embryos after fresh embryo transfer or blastocysts from freeze-all cycles. All embryos were transferred 2- 4h after thawing.

Participants/materials, setting, methods
We compared reproductive outcomes of slow-growing embryos frozen on D5 versus (n = 442) slow-growing embryos frozen on D6 (n = 379). D5 group consisted in embryos graded 0, 1, 2 of Gardner scale and frozen on D5. Similarly, D6 group consisted in embryos graded 3, 4, 5 of Gardner scale (blastocyst stage) and frozen on D6. Differences in pregnancy rates between study groups were compared using a Chi2 test. A p-value &lt;0.05 was considered statistically significant.

Main results and the role of chance
Baseline characteristics were comparable between study groups. Overall, mean age of the woman was 42.3±5.4 years old; donor sperm was used in 25% of cycles, and it was frozen in 73.2% of cycles. Pregnancy rates were significantly higher when transferring slow D5 embryos compared to D6 for all the pregnancy outcomes analyzed: biochemical pregnancy rate was 27.7% vs 20.2%, p &lt; 0.016; clinical pregnancy rate was 17.5% vs 10.2%, p &lt; 0.004); ongoing pregnancy rate was: 15.7% vs 7.8% (p &lt; 0.001); live birth rate was: 15.4% vs 7.5%, (p &lt; 0.001). These results suggest that when embryos exhibit a slow development behavior (not reaching full blastocysts at D5), waiting until D6 for blastulation and expansion does not improve clinical outcomes. Vitrification at D5 will should the preferred option in cases where the oocyte is assumed of high quality

Limitations, reasons for caution
The retrospective design of the study is its main limitation. Also, morphology as sole selection criterion for transfer. However, blastocyst morphology is a very good predictor of implantation and pregnancy, and a good indicator of the embryo’s chromosomal status (higher euploidy rate in higher morphological quality blastocysts).
Wider implications of the findings: These results can help to the standardization of laboratory protocols. As the decision of vitrifying slow developing embryos on D5 or D6 is made by the laboratory team or by the gynaecologist in agreement with the patient, having an evidence based strategy simplifies patient counselling and decision making.

Trial registration number
Not applicable