Pre-Vitrification and Post-Warming Variables of Vitrified-Warmed Blastocysts That Are Predictable for Implantation

Anette Gabrielsen, Lea Hedegaard Iversen, Jens Fedder, Tilde Veng Eskildsen, Anne Lis Englund, Stine Ravn Hansen, Philippe Pinton, Anette Gabrielsen, Lea Hedegaard Iversen, Jens Fedder, Tilde Veng Eskildsen, Anne Lis Englund, Stine Ravn Hansen, Philippe Pinton

Abstract

Human IVF embryos that are not used for fresh transfer are cryopreserved by vitrification for later embryo transfers. This study evaluates pre-vitrification and post-warming embryo characteristics that are suitable to predict the chance of clinical pregnancy in single vitrified blastocyst transfer (SVBT) cycles. In a multicenter observational trial (IMBOS trial), embryos were cultured in a time-lapse system before and after vitrification. Associations between clinical pregnancy, morphokinetic parameters, blastocyst collapse, KIDScore D5, pre-vitrification and post-warming Gardner scores, post-warming blastocyst size and re-expansion rates before SVBT were analyzed in 182 SVBTs which resulted in 89 clinical pregnancies. No association was found between clinical pregnancy after SVBT and the number of collapses or the maximal collapse size before vitrification. The multifactorial analysis of pre-vitrification Gardner scores showed a significant association with clinical pregnancy for trophectoderm grading but not for expansion/hatching status and inner cell mass grading. A significant association with clinical pregnancy was found for the time to reach a blastocyst after pronuclear fading (tB-tPNf), KIDScore D5 and post-warming size but not the rate of expansion or maximal expansion size. The selection of blastocysts for SVBT could benefit from using pre-vitrification parameters like tB-tPNf, trophectoderm grading and post-warming blastocyst size.

Keywords: blastocyst; implantation; morphokinetics; morphology; pregnancy; prognostic factors; time-lapse; transfer; vitrification; warming.

Conflict of interest statement

A.G., L.H.I., T.V.E., J.F., A.L.E. and S.R.H. report no conflicts of interest. P.P. is an employee of Ferring Pharmaceuticals and owns stocks of Takeda Pharmaceuticals.

Figures

Figure 1
Figure 1
Correlation of Gardner and Schoolcraft score before vitrification and cryotransfer with a probability of a clinical pregnancy. The probability of clinical pregnancy in relation to the Gardner and Schoolcraft grading score is shown for 182 SVBT cycles in 130 subjects: (a,b) before vitrification and cryotransfer for expansion and hatching; (c,d) for ICM; (e,f) and trophectoderm, respectively. In univariate analysis, ICM and trophectoderm were significant before vitrification and cryotransfer, as indicated in the respective graph. ICM, inner cell mass; SVBT, single vitrified blastocyst transfer.
Figure 2
Figure 2
Probability of clinical pregnancy with number of collapses and maximal collapse size before vitrification. (a) The probability of clinical pregnancy in relation to the number of collapses during culture before vitrification (from 182 SVBT cycles in 129 subjects); (b) The association between the probability of clinical pregnancy and maximal collapse size as a percentage (from 179 SVBT cycles in 128 subjects) is based on the statistical analysis with slope (black line) and p-value indicated; neither were statistically significant. SVBT, single vitrified blastocyst transfer.
Figure 3
Figure 3
Probability of clinical pregnancy with blastocyst size after warming, maximal re-expansion and exponential rate of re-expansion. (a) Data on blastocyst size after warming in µm2; (b) Maximal re-expansion size in µm2; and (c) Exponential rate of re-expansion in %. For each graph, the datasets were grouped in approximately 10 equal parts with the number of cycles indicated for each bar. In each graph, the curve shows the estimated probabilities for pregnancy based on statistical analysis with the slope gradient (regression line) and p-value indicated. SVBT, single vitrified blastocyst transfer.
Figure 4
Figure 4
The probability of clinical pregnancy with tB-tPNf during culture and the association of tB-tPNf with size after warming and the exponential rate of re-expansion. (a) Data on development from the time of pronuclear fading to time of full blastocyst (tB-tPNf) from 180 SVBT cycles in 128 subjects were grouped in approximately 10 equal parts with the number of cycles indicated for each bar. The estimated probability for clinical (vital) pregnancy is based on the statistical analysis with slope and p-value indicated. The association between (b) tB-tPNf and size after warming and (c) the exponential rate of re-expansions are shown with the corresponding p-value and slope gradient (regression line).
Figure 4
Figure 4
The probability of clinical pregnancy with tB-tPNf during culture and the association of tB-tPNf with size after warming and the exponential rate of re-expansion. (a) Data on development from the time of pronuclear fading to time of full blastocyst (tB-tPNf) from 180 SVBT cycles in 128 subjects were grouped in approximately 10 equal parts with the number of cycles indicated for each bar. The estimated probability for clinical (vital) pregnancy is based on the statistical analysis with slope and p-value indicated. The association between (b) tB-tPNf and size after warming and (c) the exponential rate of re-expansions are shown with the corresponding p-value and slope gradient (regression line).

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