Embryo morphokinetic score is associated with biomarkers of developmental competence and implantation

J Assist Reprod Genet. 2021 Jul;38(7):1737-1743. doi: 10.1007/s10815-021-02162-9. Epub 2021 Apr 5.


Purpose: To study embryo morphokinetics in relation to release in spent media of molecules with possible roles in development and implantation (miR-20a, miR-30c, and sHLA-G).

Methods: Data were obtained from embryos generated in standard IVF and ICSI cycles. The Eeva system was used for embryo assessment, based on early morphokinetic parameters and producing a score (1-5, best-worst) corresponding to higher/medium/lower chances of development to blastocyst. miRNAs - mm miR-20a-5p and miR-30c-5p - and sHLA-G were quantified in 25 μl of spent blastocyst media (SBM) collected before vitrification or transfer. Statistical analyses were performed applying Kolmogorov-Smirnov, Shapiro-Wilk, and Spearman's correlation coefficient tests, where appropriate.

Results: SBM were collected from a total of 172 viable blastocysts. Their analysis showed that concentration of miR-20a was progressively lower as Eeva score increased and probability of development to blastocyst decreased (P = 0.016). The opposite trend was observed in the case of miR-30c, i.e., concentration was higher as score increased and chances of development to blastocyst decreased (P = 0.004). Analysis of sHLA-G revealed a negative correlation with Eeva score, i.e., levels were progressively lower as Eeva score increased and probability of development to blastocyst decreased (R = - 0.388, N = 141, P = 0.001).

Conclusion: Our data suggest that morphokinetic algorithms that predict development to blastocyst stage, in fact, also identify embryos with molecular and cellular profiles more consistent with developmental functions.

Keywords: Blastocyst; Embryo; Implantation; In vitro fertilization; miRNA; sHLA-G.

MeSH terms

  • Adult
  • Biomarkers / analysis
  • Blastocyst / cytology
  • Blastocyst / physiology*
  • Bone Substitutes
  • Culture Media / analysis
  • Embryo Culture Techniques / methods
  • Embryo Implantation / physiology*
  • Female
  • Fertilization in Vitro
  • Gene Expression Regulation, Developmental
  • HLA-G Antigens / analysis*
  • Humans
  • Male
  • MicroRNAs / analysis*
  • Proof of Concept Study


  • Algipore
  • Biomarkers
  • Bone Substitutes
  • Culture Media
  • HLA-G Antigens
  • MIRN20a microRNA, human
  • MIRN30C2 microRNA, human
  • MicroRNAs