title: Morphokinetic analysis and embryonic prediction for blastocyst formation through an integrated time-lapse system.
abstract: OBJECTIVE
  To describe the events associated with the blastocyst formation and implantation that occur in embryos during preimplantation development based on the largest sample size ever described with time-lapse monitoring.
  
  
  METHODS
  Observational, retrospective, single-center clinical study.
  
  
  METHODS
  University-affiliated private IVF center.
  
  
  METHODS
  A total of 7,483 zygotes from 990 first treatments of intracytoplasmic sperm injection (ICSI; 627 of oocyte donor vs. 363 autologous oocyte cycles), of which 832 blastocysts were transferred.
  
  
  METHODS
  No patient intervention. Embryos were cultured in a time-lapse monitoring system, and the embryos were transferred on day 5 after ICSI. Embryo selection was based on the multivariable model previously developed and on blastocyst morphology.
  
  
  METHODS
  Using a time-lapse system, embryo images were acquired every 15 minutes for 120 hours. Embryos cleavage time points up to the 9-cell stage (t2-t9) as well as to the morula stage (tM) and blastocyst formation (tB) were registered in hours after ICSI. Additionally, duration of the cell cycle and synchrony of the second and third cell cycles were defined. As a result, we have monitored the embryonic development of a total of 3,215 blastocysts, of which 832 were transferred. Finally, we analyzed the characteristics of embryonic development of blastocyst (phase 1) and of implanted and not implanted (phase 2) embryos as finally validated in an independent data set (phase 3).
  
  
  RESULTS
  A detailed retrospective analysis of cleavage times was made for 7,483 zygotes. We analyzed 17 parameters and found several significantly correlated with subsequent blastocyst formation and implantation. The most predictive parameters for blastocyst formation were time of morula formation, tM (81.28-96.0 hours after ICSI), and t8-t5 (≤8.78 hours) or time of transition of 5-blastomere embryos to 8-blastomere embryos with a receiver operating characteristic curve (ROC) value = 0.849 (95% confidence interval [CI], 0.835-0.854; phase 1). These parameters were less predictive of implantation, with a ROC value = 0.546 (95% CI, 0.507-0.585). We also observed that time for expansion blastocyst, tEB (107.9-112.9 hours after ICSI), and t8-t5 (≤5.67 hours after ICSI) predict blastocyst implantation, with a ROC value = 0.591 (95% CI, 0.552-0.630; phase 2). The model was validated on an independent data set and gave a ROC of 0.596 (0.526-0.666; phase 3).
  
  
  CONCLUSIONS
  The inclusion of kinetic parameters into score evaluation may improve blastocyst selection criteria and can predict blastocyst formation with high accuracy. We propose two multivariable models based on our findings to classify embryos according to their probabilities of blastocyst stage and implantation in the largest data set ever reported of human blastocysts.
authors.affiliation: Instituto Valenciano de Infertilidad, Universidad de Valencia, Valencia, Spain.
authors.forename: Yamileth
authors.initials: Y
authors.lastname: Motato
authors.affiliation: Instituto Valenciano de Infertilidad, Universidad de Valencia, Valencia, Spain.
authors.forename: María José
authors.initials: MJ
authors.lastname: de los Santos
authors.affiliation: Instituto Valenciano de Infertilidad, Universidad de Valencia, Valencia, Spain.
authors.forename: María José
authors.initials: MJ
authors.lastname: Escriba
authors.affiliation: Instituto Valenciano de Infertilidad, Universidad de Valencia, Valencia, Spain.
authors.forename: Belén Aparicio
authors.initials: BA
authors.lastname: Ruiz
authors.affiliation: Instituto Valenciano de Infertilidad, Universidad de Valencia, Valencia, Spain.
authors.forename: José
authors.initials: J
authors.lastname: Remohí
authors.affiliation: Instituto Valenciano de Infertilidad, Universidad de Valencia, Valencia, Spain. Electronic address: marcos.meseguer@ivi.es.
authors.forename: Marcos
authors.initials: M
authors.lastname: Meseguer
mesh_terms: D000465:Algorithms; D001755:Blastocyst / Q000502:physiology*; D046149:Embryo Culture Techniques; D010064:Embryo Implantation; D004624:Embryo Transfer; D005260:Female; D005298:Fertility; D006801:Humans; D007090:Image Interpretation, Computer-Assisted; D007246:Infertility / Q000175:diagnosis / Q000503:physiopathology / Q000628:therapy*; D007700:Kinetics; D009024:Morphogenesis; D015999:Multivariate Analysis; D011247:Pregnancy; D012189:Retrospective Studies; D013030:Spain; D020554:Sperm Injections, Intracytoplasmic; D059008:Time-Lapse Imaging / Q000295:instrumentation*; D016896:Treatment Outcome; D015053:Zygote / Q000502:physiology*
keywords: Embryo quality; assisted reproductive technologies; implantation; morphokinetic parameters; time lapse