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Purpose <p>To determine whether day 2 to day 3 developmental trajectories improve day 3 embryo ranking and selection compared with day 3 cell number alone, thereby offering better prognostic information for cleavage-stage embryos, including fast-cleaving embryos whose developmental potential remains uncertain.</p> Design <p>This retrospective multicenter study (2016–2024) developed a ranking framework using 27,002 embryos from one center and validated it in 1691 fresh day 3 single embryo transfers across three centers (maternal age ≤ 38&#xa0;years; embryos derived from two pronuclei, with fragmentation ≤ 10%, symmetric, and no multinucleation for confounder control). Developmental trajectories combined day 2 and day 3 cell numbers. Mixed-effects logistic models assessed trajectory–blastulation associations; post hoc comparisons consolidated trajectories into four ranking categories. Validation used trend tests between ranking levels and pregnancy rates, multivariable mixed-effects logistic model to estimate adjusted odds ratios (aORs) between ranking levels, and AUC for pregnancy prediction.</p> Results <p>Trajectory analysis reversed the advantage of day 3 8-cell over 9–16-cell embryos (30.7% vs. 24.2% high-quality blastulation; <i>P</i> &lt; 0.001), with Normal_Fast (4 → 9–16) achieving the highest rate (40.7%), significantly higher than Normal_Normal (4 → 8; 35.1%; <i>P</i> &lt; 0.001). Trajectories were consolidated into a four-category ranking: (1) Normal_Fast (4 → 9–16); (2) Normal_Normal (4 → 8); (3) Normal_Slow (4 → 6–7), Slow_Normal (&lt; 4 → 8), Fast_Normal and Fast_Fast (&gt; 4 → 8–16); and (4) all others. Ranking 1 had a higher likelihood of clinical pregnancy than ranking 2 (aOR = 2.06 [1.05–6.82]), although this validation was limited by sample size (<i>n</i> = 24). Trajectory outperformed day 3 cell number in predicting high-quality blastocyst (AUC 0.776 vs. 0.729; <i>P</i> &lt; 0.001) and clinical pregnancy (AUC original 0.619 vs. 0.596, <i>P</i> = 0.02; down-sampled 0.682 vs. 0.618, <i>P</i> = 0.016).</p> Conclusions <p>For women ≤ 38&#xa0;years undergoing fresh day 3 single embryo transfer, developmental trajectories from day 2 to day 3 improve embryo ranking and selection with other morphological features controlled (two pronuclei, fragmentation ≤ 10%, symmetry, no multinucleation). The 4 → 9–16 pathway emerges as potentially optimal, though requiring further validation. This dual-day framework treats both cell numbers as interdependent markers, where day 2 status redefines the priority of day 3 cell number, rather than serving only as a secondary criterion.</p>

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Developmental trajectories from day 2 to day 3 refine ranking and selection of cleavage-stage embryos

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2026年7月25日

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  • Wen-jie Huo,
  • Fei Peng,
  • Chen Luo,
  • Zhi-heng Chen,
  • Qiu-xia Yan,
  • Ling Sun,
  • Cai-rong Chen,
  • Song Quan,
  • Xiao-cong Wang

摘要

Purpose

To determine whether day 2 to day 3 developmental trajectories improve day 3 embryo ranking and selection compared with day 3 cell number alone, thereby offering better prognostic information for cleavage-stage embryos, including fast-cleaving embryos whose developmental potential remains uncertain.

Design

This retrospective multicenter study (2016–2024) developed a ranking framework using 27,002 embryos from one center and validated it in 1691 fresh day 3 single embryo transfers across three centers (maternal age ≤ 38 years; embryos derived from two pronuclei, with fragmentation ≤ 10%, symmetric, and no multinucleation for confounder control). Developmental trajectories combined day 2 and day 3 cell numbers. Mixed-effects logistic models assessed trajectory–blastulation associations; post hoc comparisons consolidated trajectories into four ranking categories. Validation used trend tests between ranking levels and pregnancy rates, multivariable mixed-effects logistic model to estimate adjusted odds ratios (aORs) between ranking levels, and AUC for pregnancy prediction.

Results

Trajectory analysis reversed the advantage of day 3 8-cell over 9–16-cell embryos (30.7% vs. 24.2% high-quality blastulation; P < 0.001), with Normal_Fast (4 → 9–16) achieving the highest rate (40.7%), significantly higher than Normal_Normal (4 → 8; 35.1%; P < 0.001). Trajectories were consolidated into a four-category ranking: (1) Normal_Fast (4 → 9–16); (2) Normal_Normal (4 → 8); (3) Normal_Slow (4 → 6–7), Slow_Normal (< 4 → 8), Fast_Normal and Fast_Fast (> 4 → 8–16); and (4) all others. Ranking 1 had a higher likelihood of clinical pregnancy than ranking 2 (aOR = 2.06 [1.05–6.82]), although this validation was limited by sample size (n = 24). Trajectory outperformed day 3 cell number in predicting high-quality blastocyst (AUC 0.776 vs. 0.729; P < 0.001) and clinical pregnancy (AUC original 0.619 vs. 0.596, P = 0.02; down-sampled 0.682 vs. 0.618, P = 0.016).

Conclusions

For women ≤ 38 years undergoing fresh day 3 single embryo transfer, developmental trajectories from day 2 to day 3 improve embryo ranking and selection with other morphological features controlled (two pronuclei, fragmentation ≤ 10%, symmetry, no multinucleation). The 4 → 9–16 pathway emerges as potentially optimal, though requiring further validation. This dual-day framework treats both cell numbers as interdependent markers, where day 2 status redefines the priority of day 3 cell number, rather than serving only as a secondary criterion.