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Updated: Aug 16, 2025

Human Blastocyst Biopsy and Vitrification
Published on: July 26, 2019
Post-Thaw Day 5 Blastocyst Culture Time Prior to Transfer Does Not Affect Assisted Reproduction Technology (ART)
Marta Ciaffaglione1, Marco Reschini1, Martina Balli1
1Infertility Unit, Fondazione IRCCS Ca' Granda Ospedale Maggiore Policlinico, 20122 Milan, Italy.
Abstract:
The frozen embryo transfer (FET) technique has been progressively used more worldwide due to improved culture conditions, as well as enhanced survival rates after vitrification. However, little is known about the effect of the post-thaw blastocyst culture duration prior to transfer on live birth rate in FET cycles. In this retrospective observational study, we evaluated the influence of two distinct post-thaw blastocyst culture spans (2-4 h versus 20-22 h) on clinical pregnancy and live birth rate. A total of n = 1927 frozen-warmed cycles were included in the analysis. Among those, n = 885 warmed blastocysts were cultured for 2-4 h, and n = 1029 were kept in culture for 20-22 h prior to transfer; the remaining blastocysts did not survive the warming protocol. We observed no significant differences in live birth and clinical pregnancy rates between the two groups. The blastocyst morphological evaluation at transfer improved following the longer culture time. No differences between the two groups were found also for gestational and neonatal outcomes. This work shows that different post-thaw embryo culture timings do not negatively impact pregnancy outcomes. Overall, these results are important in the context of the embryological laboratory in order to better organize the workflow and avoid unnecessary timing-related workload.
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