Purpose
To investigate the effect of embryos developmental stage and morphological grade on the sex ratio of neonates at birth.
Methods
This was a retrospective cohort study, and including 5339 singletons born from IVF/ICSI (fresh and frozen) embryos transfer cycles between 2011 and 2022 at the Center for Reproductive Medicine of the 901st Hospital of the Joint Logistics Support Force of PLA. Only single embryo transfer (SET) or equal grade embryos transfer and resulting in a singleton gestations and live birth were included. Donors (oocytes or sperm) and preimplantation genetic testing (PGT) treatment were excluded. Demographics, cycle characteristics and assisted reproductive technology (ART) procedures were analyzed. Logistic regression was used to evaluate the percentage of males at birth for association with embryos quality and morphological parameters.
Results
Transfer with good grade embryos (GGE) significantly increased percentage of male neonates in comparing with low grade embryos (LGE) (54.2% versus 46.0%, P = 0.001). After adjustment for potential confounding variables, logistic regression indicated that when compared to GGE, LGE decreased the percentage of male neonates (odd ratio (OR) 0.741, 95% CI 0.592–0.927, P = 0.009), as well as with low grade blastocysts (LGB) was significantly associated with reduced percentage of male neonates when compared to good grade blastocysts (GGB) (OR 0.740, 95% CI 0.591–0.926, P = 0.008), however, it has not shown a relationship between cleavage stage embryos (good and low grades) transfer and sex at birth. Moreover, in SET cycles, the speed of embryonic development (7–8 cells, ≥ 9 cells and ≤ 6 cells) in cleavage stage embryos was not shown to be significantly associated with the sex at birth (P = 0.467), in addition, the percentage of male neonates was found to be significantly different from the reference category (blastocyst grading AA, P < 0.0001), only for blastocyst grading BC (OR 0.309, 95% CI 0.141–0.676, P = 0.003) and CB (OR 0.501, 95% CI 0.253–0.992, P = 0.047), compared to the reference category in blastocyst stage embryo transfer cycles. Furthermore, the data were stratified by categorical variables (blastocyst quality and developmental culture days (day 5 and day 6/7)), after pairwise comparison, the result showed that day 5 LGB was significantly lower than day 5 GGB (46.7% versus 55.6%, P = 0.013), the variations were no statistically significant differences with day 6/7 LGB and day 6/7 GGB were found. Finally, there were no statistically significant differences in the percentage of males at birth between low grade cleavage stage embryos (LGC) and good grade cleavage stage embryos (GGC) (47.5% versus 52.9%, respectively).
Conclusion
The morphological scores of cleavage stage embryos were not associated with sex ratio bias. The transfer of GGB has a higher probability of being male than LGB, particularly for day 5 GGB.