Top

Journal of Assisted Reproduction and Genetics

Published in:

01-02-2019 | Genetics

Chromosome constitution of equal-sized three-cell embryos using next-generation sequencing technology

Authors: Minyue Ma, Shihui Zhang, Chongzhao Lu, Shuling Wang, Yuanqing Yao, Hongmei Peng

Published in: Journal of Assisted Reproduction and Genetics | Issue 2/2019

Abstract

Purpose

To study the chromosome constitution of equal-sized three-cell embryo.

Methods

We determined the chromosome constitution of 105 blastomeres from 35 embryos using multiple annealing and looping-based amplification cycles (MALBAC) together with NGS sequencing technology. Chromosomal copy number variation (CNV) analysis was successfully performed in 27 embryos. We also analyzed radius, perimeter, area, and volume of each blastomere to explore the possibility of selecting the normal embryos.

Results

Majority of the embryos (77.8%, 21/27) studied were mosaic or aneuploid, and only 22.2% (6/27) had normal chromosome numbers. The aneuploid chromosomes spread across all chromosomes and the most frequent aneuploidies were for chromosomes 1, 16, and 18 followed by 13, 19, and 21. Statistical analyses showed no significant difference between euploid and aneuploid embryos regarding radius, perimeter, area, and volume of their blastomeres.

Conclusions

Our results showed that majority of the equal-sized three-cell embryos were chromosomally abnormal and could not be distinguished by morphology observation, so they should be given lower priority at selection for transfer.

Available only for authorised users

Alpha Scientists in Reproductive M, Embryology ESIGo. The Istanbul consensus workshop on embryo assessment: proceedings of an expert meeting. Hum Reprod. 2011;26:1270–83.CrossRef

Prados FJ, Debrock S, Lemmen JG, Agerholm I. The cleavage stage embryo. Hum Reprod. 2012;27(Suppl 1):i50–71.CrossRefPubMed

Hlinka D, Kalatova B, Uhrinova I, Dolinska S, Rutarova J, Rezacova J, et al. Time-lapse cleavage rating predicts human embryo viability. Physiol Res. 2012;61:513–25.PubMed

Athayde Wirka K, Chen AA, Conaghan J, Ivani K, Gvakharia M, Behr B, et al. Atypical embryo phenotypes identified by time-lapse microscopy: high prevalence and association with embryo development. Fertil Steril. 2014;101:1637–48 e1–5.CrossRefPubMed

Telentschak S, Soliwoda M, Nohroudi K, Addicks K, Klinz FJ. Cytokinesis failure and successful multipolar mitoses drive aneuploidy in glioblastoma cells. Oncol Rep. 2015;33:2001–8.CrossRefPubMed

Kalatova B, Jesenska R, Hlinka D, Dudas M. Tripolar mitosis in human cells and embryos: occurrence, pathophysiology and medical implications. Acta Histochem. 2015;117:111–25.CrossRefPubMed

Chamayou S, Patrizio P, Storaci G, Tomaselli V, Alecci C, Ragolia C, et al. The use of morphokinetic parameters to select all embryos with full capacity to implant. J Assist Reprod Genet. 2013;30:703–10.CrossRefPubMedPubMedCentral

Zhan Q, Ye Z, Clarke R, Rosenwaks Z, Zaninovic N. Direct unequal cleavages: embryo developmental competence, genetic constitution and clinical outcome. PLoS One. 2016;11:e0166398.CrossRefPubMedPubMedCentral

Hardarson T, Selleskog U, Reismer E, Wigander A, Wennerstrom S, Westin C, et al. Zygotes cleaving directly into more than two cells after 25–27 h in culture are predominantly chromosomally abnormal. Hum Reprod. 2006;21:1.CrossRef

10.

Fiorentino F, Bono S, Biricik A, Nuccitelli A, Cotroneo E, Cottone G, et al. Application of next-generation sequencing technology for comprehensive aneuploidy screening of blastocysts in clinical preimplantation genetic screening cycles. Hum Reprod. 2014;29:2802–13.CrossRefPubMed

11.

Yan L, Huang L, Xu L, Huang J, Ma F, Zhu X, et al. Live births after simultaneous avoidance of monogenic diseases and chromosome abnormality by next-generation sequencing with linkage analyses. Proc Natl Acad Sci U S A. 2015;112:15964–9.CrossRefPubMedPubMedCentral

12.

Vanneste E, Voet T, Le Caignec C, Ampe M, Konings P, Melotte C, et al. Chromosome instability is common in human cleavage-stage embryos. Nat Med. 2009;15:577–83.CrossRefPubMed

13.

Lu Y, Peng H, Jin Z, Cheng J, Wang S, Ma M, et al. Preimplantation genetic diagnosis for a Chinese family with autosomal recessive Meckel-Gruber syndrome type 3 (MKS3). PLoS One. 2013;8:e73245.CrossRefPubMedPubMedCentral

14.

Zong C, Lu S, Chapman AR, Xie XS. Genome-wide detection of single-nucleotide and copy-number variations of a single human cell. Science. 2012;338:1622–6.CrossRefPubMedPubMedCentral

15.

Chan KC, Jiang P, Chan CW, Sun K, Wong J, Hui EP, et al. Noninvasive detection of cancer-associated genome-wide hypomethylation and copy number aberrations by plasma DNA bisulfite sequencing. Proc Natl Acad Sci U S A. 2013;110:18761–8.CrossRefPubMedPubMedCentral

16.

Kola I, Trounson A, Dawson G, Rogers P. Tripronuclear human oocytes: altered cleavage patterns and subsequent karyotypic analysis of embryos. Biol Reprod. 1987;37:395–401.CrossRefPubMed

17.

Meseguer M, Herrero J, Tejera A, Hilligsoe KM, Ramsing NB, Remohi J. The use of morphokinetics as a predictor of embryo implantation. Hum Reprod. 2011;26:2658–71.CrossRefPubMed

18.

Rubio I, Kuhlmann R, Agerholm I, Kirk J, Herrero J, Escriba MJ, et al. Limited implantation success of direct-cleaved human zygotes: a time-lapse study. Fertil Steril. 2012;98:1458–63.CrossRefPubMed

19.

Gleicher N, Vidali A, Braverman J, Kushnir VA, Barad DH, Hudson C, et al. Accuracy of preimplantation genetic screening (PGS) is compromised by degree of mosaicism of human embryos. Reprod Biol Endocrinol. 2016;14:54.CrossRefPubMedPubMedCentral

20.

Liu J, Wang W, Sun X, Liu L, Jin H, Li M, et al. DNA microarray reveals that high proportions of human blastocysts from women of advanced maternal age are aneuploid and mosaic. Biol Reprod. 2012;87:148.CrossRefPubMed

21.

Baart EB, Martini E, Van Den Berg I, Macklon NS, Galjaard RJ, Fauser BC, et al. Preimplantation genetic screening reveals a high incidence of aneuploidy and mosaicism in embryos from young women undergoing IVF. Hum Reprod. 2006;21:223–33.CrossRefPubMed

22.

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

23.

Chow JF, Yeung WS, Lau EY, Lee VC, Ng EH, Ho PC. Array comparative genomic hybridization analyses of all blastomeres of a cohort of embryos from young IVF patients revealed significant contribution of mitotic errors to embryo mosaicism at the cleavage stage. Reprod Biol Endocrinol. 2014;12:105.CrossRefPubMedPubMedCentral

24.

Wells D, Delhanty JD. Comprehensive chromosomal analysis of human preimplantation embryos using whole genome amplification and single cell comparative genomic hybridization. Mol Hum Reprod. 2000;6:1055–62.CrossRefPubMed

25.

Fragouli E, Alfarawati S, Spath K, Babariya D, Tarozzi N, Borini A, et al. Analysis of implantation and ongoing pregnancy rates following the transfer of mosaic diploid-aneuploid blastocysts. Hum Genet. 2017;136:805–19.CrossRefPubMed

26.

Palermo G, Munne S, Cohen J. The human zygote inherits its mitotic potential from the male gamete. Hum Reprod. 1994;9:1220–5.CrossRefPubMed

27.

Sathananthan AH, Tarin JJ, Gianaroli L, Ng SC, Dharmawardena V, Magli MC, et al. Development of the human dispermic embryo. Hum Reprod Update. 1999;5:553–60.CrossRefPubMed

28.

Gisselsson D, Jin Y, Lindgren D, Persson J, Gisselsson L, Hanks S, et al. Generation of trisomies in cancer cells by multipolar mitosis and incomplete cytokinesis. Proc Natl Acad Sci U S A. 2010;107:20489–93.CrossRefPubMedPubMedCentral

29.

Chatzimeletiou K, Morrison EE, Prapas N, Prapas Y, Handyside AH. Spindle abnormalities in normally developing and arrested human preimplantation embryos in vitro identified by confocal laser scanning microscopy. Hum Reprod. 2005;20:672–82.CrossRefPubMed

Title: Chromosome constitution of equal-sized three-cell embryos using next-generation sequencing technology
Authors: Minyue Ma
Shihui Zhang
Chongzhao Lu
Shuling Wang
Yuanqing Yao
Hongmei Peng
Publication date: 01-02-2019
Publisher: Springer US
Published in: Journal of Assisted Reproduction and Genetics / Issue 2/2019
Print ISSN: 1058-0468
Electronic ISSN: 1573-7330
DOI: https://doi.org/10.1007/s10815-018-1362-7

At a glance: The ONWARDS insulin icodec trials

Springer Medicine

Chromosome constitution of equal-sized three-cell embryos using next-generation sequencing technology

Abstract

Purpose

Methods

Results

Conclusions

At a glance: The ONWARDS insulin icodec trials

Springer Medicine

Abstract

Purpose

Methods

Results

Conclusions

Please log in to get access to this content

Other articles of this Issue 2/2019

Extracellular vesicle mediated embryo-endometrial cross talk during implantation and in pregnancy

Treatment of female rhesus macaques with a somatostatin receptor antagonist that increases oocyte fertilization rates without affecting post-fertilization development outcomes

The mitochondrial DNA content of cumulus cells may help predict embryo implantation

Chemotherapy-related damage to ovarian reserve in childhood cancer survivors: interpreting the evidence

Detailed endometrial immune assessment of both normal and adverse reproductive outcome populations

Limited relationships between reactive oxygen species levels in culture media and zygote and embryo development