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Journal of Assisted Reproduction and Genetics

24-04-2025 | ICSI | Review

Scoping review on the embryonic developmental potential of tripronuclear zygotes and their use for clinical and research purposes

Authors: Anne Mayeur, Khadija Ouaziz, Brian Sperelakis-Beedham, Paula Rubens, Nadine Gigarel, Julie Steffann, Nelly Frydman

Published in: Journal of Assisted Reproduction and Genetics

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Abstract

We reviewed the published literature on human 3PN zygotes, focusing on their developmental potential, genetic status, and possible correction, either spontaneous (autocorrection) or through micromanipulation. A total of 29 articles published between 2000 and 2023 were deemed eligible for this scoping review. Significant differences exist between the developmental potential of bipronucleated zygotes and 3PN zygotes, with the latter showing a lower blastulation rate and altered cleavage patterns, and this is related to the fertilization method used. 3PN embryos obtained by conventional IVF are most often diandric, and arrest their development more quickly with a lower blastulation rate, compared to those obtained by ICSI. This result may be linked to the number of centrioles establishing the first cytokinesis. Autocorrection can occur in 3PN zygotes, resulting in diploid embryos, and there have been reports of live births following the transfer of 3PN embryos. Induced diploidization through pronuclear removal by micromanipulation is technically feasible and associated with good survival rates. Because tripronuclear embryos are destined to be discarded, they can serve as a source of additional embryos for research purposes without major ethical concerns. Furthermore, the transfer of 3PN embryos, corrected either spontaneously or by micromanipulation, holds the potential to increase the pool of embryos eligible for transfer in IVF centers. However, this cannot be considered without genetic screening aiming to evaluate diploidy, euploidy, and heteroparental inheritance, and no studies have yet evaluated these three genetic aspects comprehensively. Further research is essential to fully understand the physiology and potential of corrected 3PN zygotes in the fields of assisted reproduction and research.
Literature
1.
Nigg EA, Raff JW. Centrioles, centrosomes, and cilia in health and disease. Cell. 2009;139:663–78.PubMedCrossRef
2.
Barrett SL, Albertini DF. Allocation of gamma-tubulin between oocyte cortex and meiotic spindle influences asymmetric cytokinesis in the mouse oocyte. Biol Reprod. 2007;76:949–57.PubMedCrossRef
3.
Sathananthan AH, Selvaraj K, Girijashankar ML, Ganesh V, Selvaraj P, Trounson AO. From oogonia to mature oocytes: inactivation of the maternal centrosome in humans. Microsc Res Tech. 2006;69:396–407.PubMedCrossRef
4.
5.
Sathananthan AH, Kola I, Osborne J, Trounson A, Ng SC, Bongso A, et al. Centrioles in the beginning of human development. Proc Natl Acad Sci U S A. 1991;88:4806–10.PubMedPubMedCentralCrossRef
6.
Schatten G. The centrosome and its mode of inheritance: the reduction of the centrosome during gametogenesis and its restoration during fertilization. Dev Biol. 1994;165:299–335.PubMedCrossRef
7.
Cavazza T, Takeda Y, Politi AZ, Aushev M, Aldag P, Baker C, et al. Parental genome unification is highly error-prone in mammalian embryos. Cell. 2021;184:2860–2877.e22.PubMedPubMedCentralCrossRef
8.
De Coster T, Masset H, Tšuiko O, Catteeuw M, Zhao Y, Dierckxsens N, et al. Parental genomes segregate into distinct blastomeres during multipolar zygotic divisions leading to mixoploid and chimeric blastocysts. Genome Biol. 2022;23:201.PubMedPubMedCentralCrossRef
9.
Tarozzi N, Nadalini M, Coticchio G, Zacà C, Lagalla C, Borini A. The paternal toolbox for embryo development and health. Mol Hum Reprod. 2021;27:gaab042.
10.
Coticchio G, Cimadomo D, Cermisoni GC, Rienzi L, Papaleo E, Ubaldi FM, et al. The first mitotic division: a perilous bridge connecting the zygote and the early embryo. Hum Reprod. 2023;38:1019–27.PubMedPubMedCentralCrossRef
11.
Coticchio G, Mignini Renzini M, Novara PV, Lain M, De Ponti E, Turchi D, et al. Focused time-lapse analysis reveals novel aspects of human fertilization and suggests new parameters of embryo viability. Hum Reprod. 2018;33:23–31.PubMedCrossRef
12.
Pieters MH, Dumoulin JC, Ignoul-Vanvuchelen RC, Bras M, Evers JL, Geraedts JP. Triploidy after in vitro fertilization: cytogenetic analysis of human zygotes and embryos. J Assist Reprod Genet. 1992;9:68–76.PubMedCrossRef
13.
Porter R, Han T, Tucker MJ, Graham J, Liebermann J, Sills ES. Estimation of second polar body retention rate after conventional insemination and intracytoplasmic sperm injection: in vitro observations from more than 5000 human oocytes. J Assist Reprod Genet. 2003;20:371–6.PubMedPubMedCentralCrossRef
14.
Macas E, Imthurn B, Roselli M, Keller PJ. Chromosome analysis of single- and multipronucleated human zygotes proceeded after the intracytoplasmic sperm injection procedure. J Assist Reprod Genet. 1996;13:345–50.PubMedCrossRef
15.
Staessen C, Van Steirteghem AC. The chromosomal constitution of embryos developing from abnormally fertilized oocytes after intracytoplasmic sperm injection and conventional in-vitro fertilization. Hum Reprod. 1997;12:321–7.PubMedCrossRef
16.
17.
Capalbo A, Treff N, Cimadomo D, Tao X, Ferrero S, Vaiarelli A, et al. Abnormally fertilized oocytes can result in healthy live births: improved genetic technologies for preimplantation genetic testing can be used to rescue viable embryos in in vitro fertilization cycles. Fertil Steril. 2017;108:1007–1015.e3.PubMedCrossRef
18.
Yalçınkaya E, Özay A, Ergin EG, Öztel Z, Özörnek H. Live birth after transfer of a tripronuclear embryo: an intracytoplasmic sperm injection as a combination of microarray and time-lapse technology. Turk J Obstet Gynecol. 2016;13:95–8.PubMedPubMedCentralCrossRef
19.
Rawlins RG, Binor Z, Radwanska E, Dmowski WP. Microsurgical enucleation of tripronuclear human zygotes. Fertil Steril. 1988;50:266–72.PubMedCrossRef
20.
Gordon JW, Grunfeld L, Garrisi GJ, Navot D, Laufer N. Successful microsurgical removal of a pronucleus from tripronuclear human zygotes. Fertil Steril. 1989;52:367–72.PubMedCrossRef
21.
Rosenbusch BE. Mechanisms giving rise to triploid zygotes during assisted reproduction. Fertil Steril. 2008;90:49–55.PubMedCrossRef
22.
Ezoe K, Takahashi T, Shimazaki K, Miki T, Tanimura Y, Amagai A, et al. Human 1PN and 3PN zygotes recapitulate all morphokinetic events of normal fertilization but reveal novel developmental errors. Hum Reprod. 2022;37:2307–19.PubMedCrossRef
23.
Escribá M-J, Martín J, Rubio C, Valbuena D, Remohí J, Pellicer A, et al. Heteroparental blastocyst production from microsurgically corrected tripronucleated human embryos. Fertil Steril. 2006;86:1601–7.PubMedCrossRef
24.
Joergensen MW, Agerholm I, Hindkjaer J, Bolund L, Sunde L, Ingerslev HJ, et al. Altered cleavage patterns in human tripronuclear embryos and their association to fertilization method: a time-lapse study. J Assist Reprod Genet. 2014;31:435–42.PubMedPubMedCentralCrossRef
25.
Jin H, Dai S, Song W, Yao G, Shi S, Sun Y. Embryo developmental potential of microsurgically corrected human three-pronuclear zygotes. Syst Biol Reprod Med. 2015;61:96–102.PubMedCrossRef
26.
Liao H-Q, OuYang Q, Zhang S-P, Cheng D-H, Lu G-X, Lin G. Pronuclear removal of tripronuclear zygotes can establish heteroparental normal karyotypic human embryonic stem cells. J Assist Reprod Genet. 2016;33:255–63.PubMedPubMedCentralCrossRef
27.
Grau N, Escrich L, Martín J, Rubio C, Pellicer A, Escribá M-J. Self-correction in tripronucleated human embryos. Fertil Steril. 2011;96:951–6.PubMedCrossRef
28.
Lammers J, Splingart C, Barrière P, Fréour T. Morphokinetic parameters of ICSI tripronucleated embryos observed using time lapse. Reprod Biomed Online. 2014;28:658–60.PubMedCrossRef
29.
Grau N, Escrich L, Galiana Y, Meseguer M, García-Herrero S, Remohí J, et al. Morphokinetics as a predictor of self-correction to diploidy in tripronucleated intracytoplasmic sperm injection-derived human embryos. Fertil Steril. 2015;104:728–35.PubMedCrossRef
30.
Takahashi H, Hirata R, Otsuki J, Habara T, Hayashi N. Are tri-pronuclear embryos that show two normal-sized pronuclei and additional smaller pronuclei useful for embryo transfer? Reprod Med Biol. 2022;21: e12462.PubMedPubMedCentralCrossRef
31.
Chen Z, Yan J, Feng HL. Aneuploid analysis of tripronuclear zygotes derived from in vitro fertilization and intracytoplasmic sperm injection in humans. Fertil Steril. 2005;83:1845–8.PubMedCrossRef
32.
Joergensen MW, Labouriau R, Hindkjaer J, Stougaard M, Kolevraa S, Bolund L, et al. The parental origin correlates with the karyotype of human embryos developing from tripronuclear zygotes. Clin Exp Reprod Med. 2015;42:14–21.PubMedPubMedCentralCrossRef
33.
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.PubMedCrossRef
34.
Yao G, Xu J, Xin Z, Niu W, Shi S, Jin H, et al. Developmental potential of clinically discarded human embryos and associated chromosomal analysis. Sci Rep. 2016;6:23995.PubMedPubMedCentralCrossRef
35.
Mutia K, Wiweko B, Iffanolida PA, Febri RR, Muna N, Riayati O, et al. The frequency of chromosomal euploidy among 3PN embryos. J Reprod Infertil. 2019;20:127–31.PubMedPubMedCentral
36.
Golubovsky M. Paternal familial twinning: hypothesis and genetic/medical implications. Twin Res. 2002;5:75–86.PubMedCrossRef
37.
Gabbett MT, Laporte J, Sekar R, Nandini A, McGrath P, Sapkota Y, et al. Molecular support for heterogonesis resulting in sesquizygotic twinning. N Engl J Med. 2019;380:842–9.PubMedCrossRef
38.
Souter VL, Parisi MA, Nyholt DR, Kapur RP, Henders AK, Opheim KE, et al. A case of true hermaphroditism reveals an unusual mechanism of twinning. Hum Genet. 2007;121:179–85.PubMedCrossRef
39.
Gu Y-F, Lin G, Lu C-F, Lu G-X. Analysis of the first mitotic spindles in human in vitro fertilized tripronuclear zygotes after pronuclear removal. Reprod Biomed Online. 2009;19:745–54.PubMedCrossRef
40.
Chen X, Fan Y, Long X, Sun X. Similar DNA methylation and histone H3 lysine 9 dimethylation patterns in tripronuclear and corrected bipronuclear human zygotes. J Reprod Dev. 2010;56:324–9.PubMedCrossRef
41.
Fan Y, Li R, Huang J, Zhao H-C, Ding T, Sun X, et al. Improved efficiency of microsurgical enucleated tripronuclear zygotes development and embryonic stem cell derivation by supplementing epidermal growth factor, brain-derived neurotrophic factor, and insulin-like growth factor-1. Stem Cells Dev. 2014;23:563–75.PubMedCrossRef
42.
Zhu H-Y, Kang X-J, Jin L, Zhang P-Y, Wu H, Tan T, et al. Histone demethylase KDM4A overexpression improved the efficiency of corrected human tripronuclear zygote development. Mol Hum Reprod. 2021;27:gaab012.
43.
Kattera S, Chen C. Normal birth after microsurgical enucleation of tripronuclear human zygotes: case report. Hum Reprod. 2003;18:1319–22.PubMedCrossRef
44.
Rungsiwiwut R, Numchaisrika P, Ahnonkitpanit V, Virutamasen P, Pruksananonda K. Triploid human embryonic stem cells derived from tripronuclear zygotes displayed pluripotency and trophoblast differentiation ability similar to the diploid human embryonic stem cells. J Reprod Dev. 2016;62:167–76.PubMedPubMedCentralCrossRef
45.
Huan Q, Gao X, Wang Y, Shen Y, Ma W, Chen Z-J. Comparative evaluation of human embryonic stem cell lines derived from zygotes with normal and abnormal pronuclei. Dev Dyn. 2010;239:425–38.PubMedCrossRef
46.
Sun X, Long X, Yin Y, Jiang Y, Chen X, Liu W, et al. Similar biological characteristics of human embryonic stem cell lines with normal and abnormal karyotypes. Hum Reprod. 2008;23:2185–93.PubMedPubMedCentralCrossRef
47.
Chen X, Luo Y, Fan Y, Yue L, Wu X, Chen Y, et al. Triploid and diploid embryonic stem cell lines derived from tripronuclear human zygotes. J Assist Reprod Genet. 2012;29:713–21.PubMedPubMedCentralCrossRef
48.
49.
Jiang C, Cai L, Huang B, Dong J, Chen A, Ning S, et al. Normal human embryonic stem cell lines were derived from microsurgical enucleated tripronuclear zygotes. J Cell Biochem. 2013;114:2016–23.PubMedCrossRef
50.
Fan Y, Li R, Huang J, Yu Y, Qiao J. Diploid, but not haploid, human embryonic stem cells can be derived from microsurgically repaired tripronuclear human zygotes. Cell Cycle. 2013;12:302–11.PubMedPubMedCentralCrossRef
51.
Liang P, Xu Y, Zhang X, Ding C, Huang R, Zhang Z, et al. CRISPR/Cas9-mediated gene editing in human tripronuclear zygotes. Protein Cell. 2015;6:363–72.PubMedPubMedCentralCrossRef
52.
Kang E, Wu J, Gutierrez NM, Koski A, Tippner-Hedges R, Agaronyan K, et al. Mitochondrial replacement in human oocytes carrying pathogenic mitochondrial DNA mutations. Nature. 2016;540:270–5.PubMedCrossRef
53.
Craven L, Tuppen HA, Greggains GD, Harbottle SJ, Murphy JL, Cree LM, et al. Pronuclear transfer in human embryos to prevent transmission of mitochondrial DNA disease. Nature. 2010;465:82–5.PubMedPubMedCentralCrossRef
54.
Golubovsky MD. Postzygotic diploidization of triploids as a source of unusual cases of mosaicism, chimerism and twinning. Hum Reprod. 2003;18:236–42.PubMedCrossRef
Metadata
Title
Scoping review on the embryonic developmental potential of tripronuclear zygotes and their use for clinical and research purposes
Authors
Anne Mayeur
Khadija Ouaziz
Brian Sperelakis-Beedham
Paula Rubens
Nadine Gigarel
Julie Steffann
Nelly Frydman
Publication date
24-04-2025
Publisher
Springer US
Published in
Journal of Assisted Reproduction and Genetics
Print ISSN: 1058-0468
Electronic ISSN: 1573-7330
DOI
https://doi.org/10.1007/s10815-025-03467-9

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