Association between growth dynamics, morphological parameters, the chromosomal status of the blastocysts, and clinical outcomes in IVF PGS cycles with single embryo transfer



The purpose of the present study is to examine interconnection between speed of embryo development, the genetic status of the blastocysts, and clinical outcomes in IVF preimplantation genetic screening (PGS) cycles with single embryo transfer (SET).


The retrospective comparative study has been performed between January 2013 and January 2016. Seven hundred thirty-seven cycles of IVF treatment with PGS, followed by 503 SETs, were included in the study. Normally fertilized oocytes were hatched on day 3, were cultured to the blastocyst stage, and were biopsied only when at least three to seven cells were herniating from zona pellucida on the morning of day 5 (≤118 h) or day 6 (≥139 h). A total of 3705 embryos were analyzed for euploidy rates and blastocyst morphology. All embryos were vitrified after the biopsy, and selected embryos were subsequently thawed for a hormone replacement frozen embryo transfer cycle.


The euploidy rate was significantly higher among embryos biopsied on day 5 versus day 6: 59.44 ± 4.1 and 48.19 ± 3.8, respectively, p < 0.05. The difference in euploidy rates between embryos biopsied on day 5 versus day 6 in matched age groups increased from 5.83 to 25.46% with advancing maternal age. Our data demonstrated no statistically significant difference in euploidy rates between good-quality embryos biopsied on day 5 in the group of patients <38 years old and embryos in PGS cycles using donor oocytes: 71.12% (336/472) and 75.68% (221/292), respectively, p = 0.174, χ 2 = 1.848. In 270 out of 503 SETs, transferred embryos were biopsied on day 5 (ongoing pregnancy rate was 64.6% in a group of patients <38 years old, and in a group of patients ≥38 years old, ongoing PR was 64.2%). In 233 out of 503 cycles, transferred embryos were biopsied on day 6 (ongoing PR was 46.6% in a group of patients <38 years old, and in a group of patients ≥38 years old, ongoing PR was 50.8%). In all study groups, the ongoing pregnancy rate was higher when the transferred embryo was available for biopsy on day 5.


Good- and fair-quality embryos available for biopsy on day 5 have higher euploidy rates and have a higher chance to result in an ongoing pregnancy. Euploidy rate has significant variations within the same age group depending on the morphology of the blastocysts.

This is a preview of subscription content, log in to check access.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5


  1. 1.

    Munné S, Wells D, Cohen J. Technology requirements for preimplantation genetic diagnosis to improve assisted reproduction outcomes. Fertil Steril. 2010;94(2):408–30.

    Article  PubMed  Google Scholar 

  2. 2.

    Aoki F, Worrad DM, Schultz RM. Regulation of transcriptional activity during the first and second cell cycles in the preimplantation mouse embryo. Dev Biol. 1997;181(2):296–307.

    CAS  Article  PubMed  Google Scholar 

  3. 3.

    Edwards RG, Beard HK. Oocyte polarity and cell determination in early mammalian embryos. Mol Hum Reprod. 1997;3(10):863–905.

    CAS  Article  PubMed  Google Scholar 

  4. 4.

    Gandolfi TALB, Gandolfi F. The maternal legacy to the embryo: cytoplasmic components and their effects on early development. Theriogenology. 2001;55(6):1255–76.

    CAS  Article  PubMed  Google Scholar 

  5. 5.

    Kuliev A, Verlinsky Y. Preimplantation genetic diagnosis: technological advances to improve accuracy and range of applications. Reprod BioMed Online. 2008;16(4):532–8.

    Article  PubMed  Google Scholar 

  6. 6.

    Nothias J-Y, et al. Regulation of gene expression at the beginning of mammalian development. J Biol Chem. 1995;270(38):22077–80.

    CAS  Article  PubMed  Google Scholar 

  7. 7.

    Noli L, et al. Developmental clock compromises human twin model created by embryo splitting. Hum Reprod. 2015;30(12):2774–84.

    PubMed  Google Scholar 

  8. 8.

    Magli MC, Gianaroli L, Ferraretti AP. Chromosomal abnormalities in embryos. Mol Cell Endocrinol. 2001;183:S29–34.

    CAS  Article  PubMed  Google Scholar 

  9. 9.

    Munne S, et al. Embryo morphology, developmental rates, and maternal age are correlated with chromosome abnormalities. Fertil Steril. 1995;64(2):382–91.

    CAS  Article  PubMed  Google Scholar 

  10. 10.

    Alfarawati S, Fragouli E, Colls P, Stevens J, Gutiérrez-Mateo C, Schoolcraft WB, Katz-Jaffe MG, Wells D. The relationship between blastocyst morphology, chromosomal abnormality, and embryo gender. Fertil Steril. 2011;95(2):520–4.

    Article  PubMed  Google Scholar 

  11. 11.

    Capalbo, Antonio, et al. “Correlation between standard blastocyst morphology, euploidy and implantation: an observational study in two centers involving 956 screened blastocysts”. Human Reproduction. 2014: deu033.

  12. 12.

    Barrenetxea G, et al. Blastocyst culture after repeated failure of cleavage-stage embryo transfers: a comparison of day 5 and day 6 transfers. Fertil Steril. 2005;83(1):49–53.

    Article  PubMed  Google Scholar 

  13. 13.

    Shapiro BS, et al. A comparison of day 5 and day 6 blastocyst transfers. Fertil Steril. 2001;75(6):1126–30.

    CAS  Article  PubMed  Google Scholar 

  14. 14.

    Barash OO, Ivani KA, Huen N, Lefko SC, Mackenzie CF, Weckstein LN. Association between blastocyst morphology and euploidy rates in different age groups analyzed by aCGH and SNP PGD. In Human Reproduction 2015 Jun 1 (vol. 30, pp. 379–379). Oxford, England: Oxford University Press; 2015.

  15. 15.

    Hardy K, Handyside AH, Winston RM. The human blastocyst: cell number, death and allocation during late preimplantation development in vitro. Development. 1989;107(3):597–604.

    CAS  PubMed  Google Scholar 

  16. 16.

    Jones GM, et al. Evolution of a culture protocol for successful blastocyst development and pregnancy. Hum Reprod. 1998;13(1):169–77.

    CAS  Article  PubMed  Google Scholar 

  17. 17.

    Hewitson L, et al. Unique checkpoints during the first cell cycle of fertilization after intracytoplasmic sperm injection in rhesus monkeys. Nat Med. 1999;5(4):431–3.

    CAS  Article  PubMed  Google Scholar 

  18. 18.

    Wang WH, Sun QY. Meiotic spindle, spindle checkpoint and embryonic aneuploidy. Frontiers in bioscience: a journal and virtual library. 2005;11:620–36.

    Article  Google Scholar 

  19. 19.

    Zhou B-BS, Elledge SJ. The DNA damage response: putting checkpoints in perspective. Nature. 2000;408(6811):433–9.

    CAS  Article  PubMed  Google Scholar 

  20. 20.

    Adler A, et al. Blastocyst culture selects for euploid embryos: comparison of blastomere and trophectoderm biopsies. Reprod BioMed Online. 2014;28(4):485–91.

    Article  PubMed  Google Scholar 

  21. 21.

    Dokras A, et al. Trophectoderm biopsy in human blastocysts. Hum Reprod. 1990;5(7):821–5.

    CAS  Article  PubMed  Google Scholar 

  22. 22.

    Ottolini CS, et al. Karyomapping identifies second polar body DNA persisting to the blastocyst stage: implications for embryo biopsy. Reprod BioMed Online. 2015;31(6):776–82.

    CAS  Article  PubMed  Google Scholar 

  23. 23.

    Gardner DK, Schoolcraft WB. Culture and transfer of human blastocysts. Curr Opin Obstet Gynecol. 1999;11(3):307–11.

    CAS  Article  PubMed  Google Scholar 

  24. 24.

    Balaban B, Urman B, Sertac A, Alatas C, Aksoy S, Mercan R. Blastocyst quality affects the success of blastocyst-stage embryo transfer. Fertil Steril. 2000;74(2):282–7.

    CAS  Article  PubMed  Google Scholar 

  25. 25.

    Honnma H, Baba T, Sasaki M, Hashiba Y, Ohno H, Fukunaga T, Endo T, Saito T, Asada Y. Trophectoderm morphology significantly affects the rates of ongoing pregnancy and miscarriage in frozen-thawed single-blastocyst transfer cycle in vitro fertilization. Fertil Steril. 2012;98(2):361–7.

    Article  PubMed  Google Scholar 

  26. 26.

    Schoolcraft WB, Gardner DK, Lane M, Schlenker T, Hamilton F, Meldrum DR. Blastocyst culture and transfer: analysis of results and parameters affecting outcome in two in vitro fertilization programs. Fertil Steril. 1999;72(4):604–9.

    CAS  Article  PubMed  Google Scholar 

  27. 27.

    Fritz MA. Perspectives on the efficacy and indications for preimplantation genetic screening: where are we now? Hum Reprod. 2008;23:2617–21.

    Article  PubMed  Google Scholar 

  28. 28.

    Scott RT Jr, Ferry K, Su J, Tao X, Scott K, Treff NR. Comprehensive chromosome screening is highly predictive of the reproductive potential of human embryos: a prospective, blinded, nonselection study. Fertil Steril. 2012;97:870–5.

    Article  PubMed  Google Scholar 

  29. 29.

    Munné S, Grifo J, Wells D. Mosaicism:“survival of the fittest” versus “no embryo left behind”. Fertil Steril. 2016;105(5):1146–9.

    Article  PubMed  Google Scholar 

  30. 30.

    Scott RT, Galliano D. The challenge of embryonic mosaicism in preimplantation genetic screening. Fertil Steril. 2016;105(5):1150–2.

    Article  PubMed  Google Scholar 

  31. 31.

    Munne S, et al. Egg donor aneuploidy rates significantly differ between fertility centers. Fertil Steril. 2014;102(3):e121.

    Article  Google Scholar 

  32. 32.

    Fragouli E, Alfarawati S, Spath K, Wells D. Morphological and cytogenetic assessment of cleavage and blastocyst stage embryos. Molecular human reproduction 2013:gat073.

  33. 33.

    Minasi MG, Colasante A, Riccio T, Ruberti A, Casciani V, Scarselli F, Spinella F, Fiorentino F, Varricchio MT, Greco E. Correlation between aneuploidy, standard morphology evaluation and morphokinetic development in 1730 biopsied blastocysts: a consecutive case series study. Human Reproduction. 2016: 2.

  34. 34.

    Liu Y, Chapple V, Feenan K, Roberts P, Matson P. Time-lapse deselection model for human day 3 in vitro fertilization embryos: the combination of qualitative and quantitative measures of embryo growth. Fertil Steril. 2016;105(3):656–62.

    Article  PubMed  Google Scholar 

  35. 35.

    Petersen BM, Boel M, Montag M, Gardner DK. Development of a generally applicable morphokinetic algorithm capable of predicting the implantation potential of embryos transferred on day 3. Hum Reprod. 2016;31(10):2231–44.

    Article  PubMed  PubMed Central  Google Scholar 

  36. 36.

    Coonen E, Derhaag JG, Dumoulin JC, van Wissen LC, Bras M, Janssen M, Evers JL, Geraedts JP. Anaphase lagging mainly explains chromosomal mosaicism in human preimplantation embryos. Hum Reprod. 2004;19(2):316–24.

    Article  PubMed  Google Scholar 

  37. 37.

    Delhanty JD. The origins of genetic variation between individual human oocytes and embryos: implications for infertility. Hum Fertil. 2013;16(4):241–5.

    Article  Google Scholar 

  38. 38.

    Kovalevsky G, Carney SM, Morrison LS, Boylan CF, Neithardt AB, Feinberg RF. Should embryos developing to blastocysts on day 7 be cryopreserved and transferred: an analysis of pregnancy and implantation rates. Fertil Steril. 2013;100(4):1008–12.

    Article  PubMed  Google Scholar 

  39. 39.

    Su Y, Li JJ, Wang C, Haddad G, Wang WH. Aneuploidy analysis in day 7 human blastocysts produced by in vitro fertilization. Reprod Biol Endocrinol. 2016;14(1):1.

    Article  Google Scholar 

  40. 40.

    Harton GL, Munné S, Surrey M, Grifo J, Kaplan B, McCulloh DH, Griffin DK, Wells D, Group PP. Diminished effect of maternal age on implantation after preimplantation genetic diagnosis with array comparative genomic hybridization. Fertil Steril. 2013;100(6):1695–703.

    CAS  Article  PubMed  Google Scholar 

  41. 41.

    HFEA. (2013). Chairs Letter, Removal of the license condition relating to multiple births. Available online at

  42. 42.

    Practice Committee of Society for Assisted Reproductive Technology and Practice Committee of American Society for Reproductive Medicine. Elective single-embryo transfer. Fertil Steril. 2012;97:835–42.

    Article  Google Scholar 

  43. 43.

    Practice Committee of the Society for Assisted Reproductive Technology and Practice Committee of the American Society for Reproductive Medicine. Guidelines on number of embryos transferred. Fertil Steril. 2009;92:1518–9.

    Article  Google Scholar 

  44. 44.

    Paternot G, Devroe J, Debrock S, D’Hooghe TM, Spiessens C. Intra- and inter-observer analysis in the morphological assessment of early-stage embryos. Reprod Biol Endocrinol. 2009;7:105.

    Article  PubMed  PubMed Central  Google Scholar 

  45. 45.

    Paternot G, Wetzels AM, Thonon F, Vansteenbrugge A, Willemen D, Devroe J, et al. Intra- and interobserver analysis in the morphological assessment of early stage embryos during an IVF procedure: a multicentre study. Reprod Biol Endocrinol. 2011;9:127.

    Article  PubMed  PubMed Central  Google Scholar 

  46. 46.

    Basile N, del Carmen NM, Bronet F, Florensa M, Riqueiros M, Rodrigo L, García-Velasco J, Meseguer M. Increasing the probability of selecting chromosomally normal embryos by time-lapse morphokinetics analysis. Fertil Steril. 2014;101(3):699–704.

    Article  PubMed  Google Scholar 

  47. 47.

    Campbell A, Fishel S, Bowman N, Duffy S, Sedler M, Hickman CF. Modelling a risk classification of aneuploidy in human embryos using non-invasive morphokinetics. Reprod BioMed Online. 2013;26(5):477–85.

    Article  PubMed  Google Scholar 

  48. 48.

    Swain JE. Could time-lapse embryo imaging reduce the need for biopsy and PGS? J Assist Reprod Genet. 2013;30(8):1081–90.

    Article  PubMed  PubMed Central  Google Scholar 

Download references

Author information




Conceived and designed the study: O.B. and K.I.; coordinated data collection: S.W. and O.B.; analyzed the data: O.B., S.W., E.R., D.W., M.H., and S.R.; and drafted the manuscript: O.B, K.I., M.H., and L.W. All authors interpreted the data.

Corresponding author

Correspondence to Oleksii O. Barash.

Ethics declarations

The study protocol was approved by the Ethics Committee (Institutional Review Board) of the Reproductive Science Center of the San Francisco Bay Area and was performed according to the Declaration of Helsinki for Medical Research.

Conflict of interest

The authors declare that they have no conflict of interest.

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Barash, O.O., Ivani, K.A., Willman, S.P. et al. Association between growth dynamics, morphological parameters, the chromosomal status of the blastocysts, and clinical outcomes in IVF PGS cycles with single embryo transfer. J Assist Reprod Genet 34, 1007–1016 (2017).

Download citation


  • Preimplantation genetic screening
  • IVF
  • CCS
  • Euploidy rates
  • Blastocyst morphology
  • Blastocyst biopsy
  • Embryo development