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Relationship between embryo morphodynamic and molecular karyotype in poor prognosis patients

P-456 Relationship between embryo morphodynamic and molecular karyotype in poor prognosis patients

  1. L. Rienzi1,
  2. A. Biricik2,
  3. A. Capalbo1,
  4. S. Colamaria1,
  5. S. Bono2,
  6. L. Spizzichino2,
  7. F. Ubaldi1 and
  8. F. Fiorentino2

+ Author Affiliations

  1. 1Genera c/o Clinica Valle Giulia, Reproductive Medicine, Roma, Italy
  2. 2GENOMA, Molecular Genetics Laboratory, Roma, Italy

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Abstract

Introduction: Just recently, time lapse cinematography has been introduced in human IVF aiming at optimizing embryo assesment. Novel parameters have been proposed according to the dynamic behavior of the embryos. Some studies reported a relationship between specific timings (5-cell division, interval between 3-cell division and subsequent 4-cell division, duration of second cell cycle) and blastocyst development and implantation. The aim of our study has been to evaluate the relationship between morphodynamic timings and molecular karyotype of embryos as assessed by array comparative genomic hybridization (aCGH).

Material and Methods: The patients enrolled in this study underwent ICSI/PGS cycles due to advanced maternal age (N = 13). The mean female age was 38.2 ± 1.8 years. All the embryos selected for PGS (N = 77) were individually monitored using a tri-gas incubator with a built-in camera designed to automatically acquire images at defined time points. Images were acquired every 30 min in nine different focal planes for at least 110h for each embryo. The embryos were subjected to biopsy either at day 3 (N = 48) or at day 5 (N = 29) and subsequently to aCGH. Briefly, blastomeres/trophoectoderm cells were first lysed and DNA was amplified by whole genome amplification (WGA) using the SurePlex kit. WGA products were then processed by aCGH according to Blugnome protocol. Embryos were classified as aneuploid when abnormalities affected one or more chromosomes. Brown–Forsythe’s test was used for the analysis of homogeneity of variances while Mann–Whitney U-test was used to test differences between median values.

Results: 25 out of 77 good quality embryos selected for the biopsy procedure resulted to be euploid (32.5%). 10 embryos were transferred in 7 patients and 4 of them implanted. No significant correlation was found between embryo cleavage timings and chromosomal complement (ie. median time values of syngamy, first, second, third and fourth cleavage). However, the time interval between 3-cell stage and 4-cell stage (s2) was significantly correlated with molecular karyotype data. In particular, median s2 time value was 1.59h in aneuploid embryos vs 1.09h in euploid embryos (CI 1.16-2.01 vs 0.71-1.48 respectively, P < 0.05). Moreover, aberrant behavior after the 2-cell stage (direct division from 2-cell to 5-cell) was correlated with complex aneuploidy in all cases (5 embryos).

Conclusions: Our data suggests that specific embryo behavior at an early developmental stage may reflect chromosomal segregation fidelity. The most sensitive moment appearing to be the sincrony and regularity of the second cell division.