The International Journal of Developmental Biology

Int. J. Dev. Biol. 64: 415 - 422 (2020)

Vol 64, Issue 7-8-9

Evaluation of morphokinetic characteristics of zona pellucida free mouse pre-implantation embryos using time-lapse monitoring system

Published: 4 August 2020

Jihyun Kim1,2, Jaewang Lee2,3, Yun Jung Choi2,4, Ohchan Kwon2, Tae Bok Lee3,5 and Jin Hyun Jun*,2,3,6

1Clinical medicine division, Korea Institute of Oriental Medicine, 2Department of Biomedical Laboratory Science, Eulji University, College of Health Science, Seongnam, 3Department of Senior Healthcare, BK21 plus Program, Graduate School of Eulji University, Seongnam, 4Seoul Rachel Fertility Center, Seoul, 5Center for Medical Innovation, Seoul National University Hospital, Seoul and 6Eulji Medi-Bio Research Institute (EMBRI), Eulji University, Daejeon, Korea


The mammalian zygote cleaves and develops to blastocyst within the zona pellucida (ZP) in vivo. The presence or absence of ZP may affect the characteristics of the embryo, including blastomere alignment, cell-cell junction, and compaction. This study aimed to compare the morphokinetic characteristics of ZP intact and ZP free mouse pre-implantation embryos with time-lapse monitoring system. Mouse 2-cell embryos were collected 1.5 days post coitum (dpc), and their ZPs were removed by treatment with acid Tyrode’s solution. All embryos were cultured in vitro up to the outgrowth stage at 7.5 dpc. In this study, ZP did not influence the cumulative times from 2-cell to further stages and blastulation. Interestingly, ZP free embryos at 4-cell stage have three patterns of blastomere alignment according to the number of contact points between blastomeres. However, blastomere alignment did not lead to any differences in morphokinetic comparisons. Regardless of the presence or absence of ZP, embryos compacted after the 8-cell stage took shorter time to become blastocysts than embryos compacted pre-8-cell stage. Nevertheless, cell-cell junction proteins required for successful compaction were similarly expressed between ZP intact and ZP free embryos. ZP intact embryos compacted post-8-cell stage had higher rate of reaching blastocysts than compacted ZP intact embryos before 8-cell stage while the outgrowth/blastocyst rate was similar. In this study, the presence or absence of ZP did not influence embryonic development and expression of cell surface glycoproteins, whereas compaction timing may be one of the criteria for evaluating embryo quality. ZP free embryos may become an alternative for overcoming cases with ZP problems in a human ART program.


timing of compaction, zona pellucida, developmental competence, time-lapse monitoring system

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