Using time-lapse technology to explore vacuolization in embryos on Day 3 and Day 4

  • Jingye Zhang
  • Wanxia Zhong
  • Hui Liu
  • Haibin Zhao
  • Mei Li
  • Shuiying Ma
  • Keliang WuEmail author
Gynecologic Endocrinology and Reproductive Medicine



To investigate the occurrence and development state of embryo vacuoles between the 8-cell and morula stages, and to explore how vacuoles affected the development of embryos.


A retrospective study of a cohort of 422 patients undergoing conventional in vitro fertilization or intracytoplasmic sperm injection. With the help of time-lapse imaging, the development processes and outcomes of good quality embryos with or without vacuoles were analyzed.


Vacuole positive embryos had significantly lower blastulation rate and good quality blastulation rate than vacuole negative embryos, p < 0.05. Compared to vacuole negative embryos, the number of best and good quality blastocysts was significantly reduced, while the number of fair and discarded ones was significantly increased, p < 0.05. The average starting time of vacuolization was 73.7 ± 9.3 h after insemination. The proportion of blastomeres affected by vacuoles was associated with embryonic developmental potential.


Vacuolization on Day 3 and Day 4 was frequently observed and was detrimental to embryo development. The proportion of blastomeres affected by vacuoles may be an indicator of embryo developmental potential.


Time-lapse Vacuoles Blastulation rate Blastocyst quality Developmental potential 


Author contributions

ZJY: data management/analysis, manuscript writing/editing. ZWX: data management/analysis, manuscript writing/editing. LH: data collection, manuscript editing. ZHB: data collection, manuscript editing. LM: manuscript editing. MSY: manuscript editing. WKL: project development, manuscript writing/editing.


This study was funded by The National Key Research and Development Program of China (2017YFC1001000) and The Shanghai Commission of Science and Technology (17DZ2271100).

Compliance with ethical standards

Conflict of interest

All authors declare that they have no conflict of interest.

Ethical approval

All procedures performed in this study involving human participants were in accordance with the ethical standards of the institutional research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards. The study was approved by the Ethics Committee of the Center for Reproductive Medicine, Shandong University.

Informed consent

Informed consent was obtained from all individual participants included in this study.


  1. 1.
    Van Blerkom J (1990) Occurrence and developmental consequences of aberrant cellular organization in meiotically mature human oocytes after exogenous ovarian hyperstimulation. J Electron Microsc Tech 16(4):324–346. CrossRefPubMedGoogle Scholar
  2. 2.
    Hardy K, Warner A, Winston RM, Becker DL (1996) Expression of intercellular junctions during preimplantation development of the human embryo. Mol Hum Reprod 2(8):621–632CrossRefGoogle Scholar
  3. 3.
    Hardy K (1999) Apoptosis in the human embryo. Rev Reprod 4(3):125–134CrossRefGoogle Scholar
  4. 4.
    Ebner T, Moser M, Sommergruber M, Gaiswinkler U, Shebl O, Jesacher K, Tews G (2005) Occurrence and developmental consequences of vacuoles throughout preimplantation development. Fertil Steril 83(6):1635–1640. CrossRefPubMedGoogle Scholar
  5. 5.
    De Sutter P, Dozortsev D, Qian C, Dhont M (1996) Oocyte morphology does not correlate with fertilization rate and embryo quality after intracytoplasmic sperm injection. Hum Reprod 11(3):595–597CrossRefGoogle Scholar
  6. 6.
    Alikani M, Palermo G, Adler A, Bertoli M, Blake M, Cohen J (1995) Intracytoplasmic sperm injection in dysmorphic human oocytes. Zygote 3(4):283–288CrossRefGoogle Scholar
  7. 7.
    Ten J, Mendiola J, Vioque J, de Juan J, Bernabeu R (2007) Donor oocyte dysmorphisms and their influence on fertilization and embryo quality. Reprod Biomed Online 14(1):40–48CrossRefGoogle Scholar
  8. 8.
    Yu EJ, Ahn H, Lee JM, Jee BC, Kim SH (2015) Fertilization and embryo quality of mature oocytes with specific morphological abnormalities. Clin Exp Reprod Med 42(4):156–162. CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Meseguer M, Herrero J, Tejera A, Hilligsoe KM, Ramsing NB, Remohi J (2011) The use of morphokinetics as a predictor of embryo implantation. Hum Reprod 26(10):2658–2671. CrossRefPubMedGoogle Scholar
  10. 10.
    Kirkegaard K, Kesmodel US, Hindkjaer JJ, Ingerslev HJ (2013) Time-lapse parameters as predictors of blastocyst development and pregnancy outcome in embryos from good prognosis patients: a prospective cohort study. Hum Reprod 28(10):2643–2651. CrossRefPubMedGoogle Scholar
  11. 11.
    Desai N, Ploskonka S, Goodman LR, Austin C, Goldberg J, Falcone T (2014) Analysis of embryo morphokinetics, multinucleation and cleavage anomalies using continuous time-lapse monitoring in blastocyst transfer cycles. Reprod Biol Endocrinol 12:54. CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Basile N, Nogales Mdel C, Bronet F, Florensa M, Riqueiros M, Rodrigo L, Garcia-Velasco J, Meseguer M (2014) Increasing the probability of selecting chromosomally normal embryos by time-lapse morphokinetics analysis. Fertil Steril 101(3):699–704. CrossRefPubMedGoogle Scholar
  13. 13.
    Campbell A, Fishel S, Bowman N, Duffy S, Sedler M, Hickman CF (2013) Modelling a risk classification of aneuploidy in human embryos using non-invasive morphokinetics. Reprod Biomed Online 26(5):477–485. CrossRefPubMedGoogle Scholar
  14. 14.
    Minasi MG, Colasante A, Riccio T, Ruberti A, Casciani V, Scarselli F, Spinella F, Fiorentino F, Varricchio MT, Greco E (2016) Correlation between aneuploidy, standard morphology evaluation and morphokinetic development in 1730 biopsied blastocysts: a consecutive case series study. Hum Reprod 31(10):2245–2254. CrossRefPubMedGoogle Scholar
  15. 15.
    Del Carmen Nogales M, Bronet F, Basile N, Martinez EM, Linan A, Rodrigo L, Meseguer M (2017) Type of chromosome abnormality affects embryo morphology dynamics. Fertil Steril 107(1):229–235. CrossRefGoogle Scholar
  16. 16.
    Gardner DK, Lane M (1997) Culture and selection of viable blastocysts: a feasible proposition for human IVF? Human Reprod Update 3(4):367–382CrossRefGoogle Scholar
  17. 17.
    Alpha Scientists In Reproductive M (2012) The Alpha consensus meeting on cryopreservation key performance indicators and benchmarks: proceedings of an expert meeting. Reprod Biomed Online 25(2):146–167. CrossRefGoogle Scholar
  18. 18.
    Gardner DK, Lane M, Stevens J, Schlenker T, Schoolcraft WB (2000) Blastocyst score affects implantation and pregnancy outcome: towards a single blastocyst transfer. Fertil Steril 73(6):1155–1158CrossRefGoogle Scholar
  19. 19.
    Majumdar G, Majumdar A, Verma IC, Upadhyaya KC (2017) Relationship between morphology, euploidy and implantation potential of cleavage and blastocyst stage embryos. J Hum Reprod Sci 10(1):49–57. CrossRefPubMedPubMedCentralGoogle Scholar
  20. 20.
    Fragouli E, Wells D (2011) Aneuploidy in the human blastocyst. Cytogenet Genome Res 133(2–4):149–159. CrossRefPubMedGoogle Scholar
  21. 21.
    Angell RR, Aitken RJ, van Look PF, Lumsden MA, Templeton AA (1983) Chromosome abnormalities in human embryos after in vitro fertilization. Nature 303(5915):336–338CrossRefGoogle Scholar
  22. 22.
    Ivec M, Kovacic B, Vlaisavljevic V (2011) Prediction of human blastocyst development from morulas with delayed and/or incomplete compaction. Fertil Steril 96(6):1473–1478. CrossRefPubMedGoogle Scholar
  23. 23.
    Tao J, Tamis R, Fink K, Williams B, Nelson-White T, Craig R (2002) The neglected morula/compact stage embryo transfer. Hum Reprod 17(6):1513–1518CrossRefGoogle Scholar
  24. 24.
    Wallbutton S, Kasraie J (2010) Vacuolated oocytes: fertilization and embryonic arrest following intra-cytoplasmic sperm injection in a patient exhibiting persistent oocyte macro vacuolization–case report. J Assist Reprod Genet 27(4):183–188. CrossRefPubMedPubMedCentralGoogle Scholar
  25. 25.
    Alikani M, Cohen J, Tomkin G, Garrisi GJ, Mack C, Scott RT (1999) Human embryo fragmentation in vitro and its implications for pregnancy and implantation. Fertil Steril 71(5):836–842CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Center for Reproductive MedicineShandong University, National Research Center for Assisted Reproductive Technology and Reproductive GeneticsJinanChina
  2. 2.The Key Laboratory for Reproductive Endocrinology (Shandong University)Ministry of EducationJinanChina
  3. 3.Center for Reproductive Medicine, Ren Ji Hospital, School of MedicineShanghai Jiao Tong UniversityShanghaiChina
  4. 4.Shanghai Key Laboratory for Assisted Reproduction and Reproductive GeneticsShanghaiChina

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