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Polar body transfer restores the developmental potential of oocytes to blastocyst stage in a case of repeated embryo fragmentation

  • Shuo-Ping Zhang
  • Chang-Fu Lu
  • Fei Gong
  • Ping-Yuan Xie
  • Liang Hu
  • Shun-Ji Zhang
  • Guang-Xiu Lu
  • Ge Lin
Technological Innovations

Abstract

Purpose

We aimed to determine the developmental potential of human reconstructed oocytes after polar body genome transfer (PBT) and to report the case of a woman with multiple cycles of severe embryo fragmentation.

Methods

Fresh and cryopreserved first polar bodies (PB1s) were transferred to enucleated metaphase II oocytes (PB1T), while fresh PB2s were removed from fertilized oocytes and used instead of the female pronucleus in donor zygotes. Reconstructed oocytes underwent intracytoplasmic sperm injection (ICSI) and were cultured to blastocyst. Biopsied trophectoderm cells of PBT-derived blastocysts were screened for chromosomes by next-generation sequencing (NGS). Then, cryopreserved PB1T was carried out in one woman with a history of several cycles of extensive embryo fragmentation, and the blastocysts derived from PB1T were screened for aneuploidy but not transferred to the patient.

Results

There were no significant differences in the rates of normal fertilization and blastocyst formation between fresh and cryopreserved PB1T and control oocytes. Of the three fresh and three cryopreserved PB1T-derived blastocysts, two and one blastocysts exhibited normal diploidy respectively. In contrast, 17 PB2 transfers yielded 16 two pronuclei (2PN) zygotes with one normal and one small-sized pronucleus each and no blastocyst formation. In the female patient, 18 oocytes were inseminated by ICSI in the fourth cycle and the PB1s were biopsied. Although the embryos developed from the patient’s own oocytes showed severe fragmentation, the oocytes reconstructed after PB1T produced three chromosomally normal blastocysts.

Conclusions

Normal blastocysts can develop from human reconstructed oocytes after PB1T. The application of the first PB transfers may be beneficial to patients with a history of poor embryo development and excessive fragmentation.

Keywords

Polar body transfer Blastocyst Embryo fragmentation Assisted reproductive technique 

Notes

Acknowledgements

This study is supported by grants from the Major State Basic Research Development Program of China (No. 2012CB944901), the National Science Foundation of China (Nos. 81222007 and 81471510), and the Program for New Century Excellent Talents in University.

Compliance with ethical standards

This study was approved by the Ethics Committee of the Reproductive and Genetic Hospital of CITIC-Xiangya (reference LL-SC-SG-2015-004).

Competing interests

The authors declare that they have no competing interests.

References

  1. 1.
    Li R, Albertini DF. The road to maturation: somatic cell interaction and self-organization of the mammalian oocyte. Nat Rev Mol Cell Biol. 2013;14:141–52.CrossRefPubMedGoogle Scholar
  2. 2.
    Zamboni L. Ultrastructure of mammalian oocytes and ova. Biol Reprod Suppl. 1970;2:44–63.CrossRefPubMedGoogle Scholar
  3. 3.
    Verlinsky Y, Ginsberg N, Lifchez A, Valle J, Moise J, Strom CM. Analysis of the first polar body: preconception genetic diagnosis. Hum Reprod. 1990;5:826–9.CrossRefPubMedGoogle Scholar
  4. 4.
    Wakayama T, Hayashi Y, Ogura A. Participation of the female pronucleus derived from the second polar body in full embryonic development of mice. J Reprod Fertil. 1997;110:263–6.CrossRefPubMedGoogle Scholar
  5. 5.
    Wakayama T, Yanagimachi R. The first polar body can be used for the production of normal offspring in mice. Biol Reprod. 1998;59:100–4.CrossRefPubMedGoogle Scholar
  6. 6.
    Wang T, Sha H, Ji D, Zhang HL, Chen D, Cao Y, et al. Polar body genome transfer for preventing the transmission of inherited mitochondrial diseases. Cell. 2014;157:1591–604.CrossRefPubMedGoogle Scholar
  7. 7.
    Wei Y, Zhang T, Wang YP, Schatten H, Sun QY. Polar bodies in assisted reproductive technology: current progress and future perspectives. Biol Reprod. 2015;92:19.CrossRefPubMedGoogle Scholar
  8. 8.
    Cohen J, Scott R, Alikani M, Schimmel T, Munné S, Levron J, et al. Ooplasmic transfer in mature human oocytes. Mol Hum Reprod. 1998;4:269–80.CrossRefPubMedGoogle Scholar
  9. 9.
    Cohen J, Scott R, Schimmel T, Levron J, Willadsen S. Birth of infant after transfer of anucleate donor oocyte cytoplasm into recipient eggs. Lancet. 1997;350:186–7.CrossRefPubMedGoogle Scholar
  10. 10.
    Dale B, Wilding M, Botta G, Rasile M, Marino M, Di Matteo L, et al. Pregnancy after cytoplasmic transfer in a couple suffering from idiopathic infertility: case report. Hum Reprod. 2001;16:1469–72.CrossRefPubMedGoogle Scholar
  11. 11.
    Prados FJ, Debrock S, Lemmen JG, Agerholm I. The cleavage stage embryo. Hum Reprod. 2012;27 Suppl 1:i50–71.CrossRefPubMedGoogle Scholar
  12. 12.
    Fujimoto VY, Browne RW, Bloom MS, Sakkas D, Alikani M. Pathogenesis, developmental consequences, and clinical correlations of human embryo fragmentation. Fertil Steril. 2011;95:1197–204.CrossRefPubMedGoogle Scholar
  13. 13.
    Meseguer M, Martínez-Conejero JA, O'Connor JE, Pellicer A, Remohí J, Garrido N. The significance of sperm DNA oxidation in embryo development and reproductive outcome in an oocyte donation program: a new model to study a male infertility prognostic factor. Fertil Steril. 2008;89:1191–9.Google Scholar
  14. 14.
    Xia P. Intracytoplasmic sperm injection: correlation of oocyte grade based on polar body, perivitelline space and cytoplasmic inclusions with fertilization rate and embryo quality. Hum Reprod. 1997;12:1750–5.CrossRefPubMedGoogle Scholar
  15. 15.
    Stensen MH, Tanbo TG, Storeng R, Åbyholm T, Fedorcsak P. Fragmentation of human cleavage-stage embryos is related to the progression through meiotic and mitotic cell cycles. Fertil Steril. 2015;103:374–81.CrossRefPubMedGoogle Scholar
  16. 16.
    Jurisicova A, Varmuza S, Casper RF. Programmed cell death and human embryo fragmentation. Mol Hum Reprod. 1996;2:93–8.CrossRefPubMedGoogle Scholar
  17. 17.
    Levy R, Benchaib M, Cordonier H, Souchier C, Guerin JF. Annexin V labelling and terminal transferase-mediated DNA end labelling (TUNEL) assay in human arrested embryos. Mol Hum Reprod. 1998;4:775–83.CrossRefPubMedGoogle Scholar
  18. 18.
    Bencomo E, Pérez R, Arteaga MF, Acosta E, Pena O, Lopez L, et al. Apoptosis of cultured granulosa-lutein cells is reduced by insulin-like growth factor I and may correlate with embryo fragmentation and pregnancy rate. Fertil Steril. 2006;85:474–80.CrossRefPubMedGoogle Scholar
  19. 19.
    Alikani M, Schimmel T, Willadsen SM. Cytoplasmic fragmentation in activated eggs occurs in the cytokinetic phase of the cell cycle, in lieu of normal cytokinesis, and in response to cytoskeletal disorder. Mol Hum Reprod. 2005;11:335–44.CrossRefPubMedGoogle Scholar
  20. 20.
    Fujimoto VY, Kane JP, Ishida BY, Bloom MS, Browne RW. High-density lipoprotein metabolism and the human embryo. Hum Reprod Update. 2010;16:20–38.CrossRefPubMedGoogle Scholar
  21. 21.
    Alikani M, Cohen J, Tomkin G, Garrisi GJ, Mack C, Scott RT. Human embryo fragmentation in vitro and its implications for pregnancy and implantation. Fertil Steril. 1999;71:836–42.CrossRefPubMedGoogle Scholar
  22. 22.
    Ebner T, Yaman C, Moser M, Sommergruber M, Pölz W, Tews G. Embryo fragmentation in vitro and its impact on treatment and pregnancy outcome. Fertil Steril. 2001;76:281–5.CrossRefPubMedGoogle Scholar
  23. 23.
    Alikani M. The origins and consequences of fragmentation in mammalian eggs and embryos. In: Elder K, Cohen J, editors. Human preimplantation embryo selection. London: Informa Healthcare; 2007. p. 51–78.CrossRefGoogle Scholar
  24. 24.
    Xu X, Duan X, Lu C, Lin G, Lu G. Dynamic distribution of NuMA and microtubules in human fetal fibroblasts, developing oocytes and somatic cell nuclear transferred embryos. Hum Reprod. 2011;26:1052–60.CrossRefPubMedGoogle Scholar
  25. 25.
    Gong F, Li X, Zhang S, Ma H, Cai S, Li J, et al. A modified ultra-long pituitary downregulation protocol improved endometrial receptivity and clinical outcome for infertile patients with polycystic ovarian syndrome. Exp Ther Med. 2015;10:1865–70.PubMedPubMedCentralGoogle Scholar
  26. 26.
    Zhang SP, Tan K, Gong F, Gu YF, Tan YQ, Lu CF, et al. Blastocysts can be rebiopsied for preimplantation genetic diagnosis and screening. Fertil Steril. 2014;102:1641–5.CrossRefPubMedGoogle Scholar
  27. 27.
    Tan YQ, Yin XY, Zhang SP, Jiang H, Tan K, Li J, et al. Clinical outcome of preimplantation genetic diagnosis and screening using next generation sequencing. Gigascience. 2014;3:30.CrossRefPubMedPubMedCentralGoogle Scholar
  28. 28.
    Hou Y, Fan W, Yan L, Li R, Lian Y, Huang J, et al. Genome analyses of single human oocytes. Cell. 2013;155:1492–506.CrossRefPubMedGoogle Scholar
  29. 29.
    Wakayama S, Hikichi T, Suetsugu R, Sakaide Y, Bui HT, Mizutani E, et al. Efficient establishment of mouse embryonic stem cell lines from single blastomeres and polar bodies. Stem Cells. 2007;25:986–93.CrossRefPubMedGoogle Scholar
  30. 30.
    Wang GJ, Yu JN, Tan XD, Zhou XL, Xu XB, Fan BQ. Injection of frozen-thawed porcine first polar bodies into enucleated oocytes results in fertilization and embryonic development. Theriogenology. 2011;75:826–31.CrossRefPubMedGoogle Scholar
  31. 31.
    Evsikov SV, Evsikov AV. Preimplantation development of manipulated mouse zygotes fused with the second polar bodies: a cytogenetic study. Int J Dev Biol. 1994;38:725–30.PubMedGoogle Scholar
  32. 32.
    VerMilyea MD, Maneck M, Yoshida N, Blochberger I, Suzuki E, Suzuki T, et al. Transcriptome asymmetry within mouse zygotes but not between early embryonic sister blastomeres. EMBO J. 2011;30:1841–51.CrossRefPubMedPubMedCentralGoogle Scholar
  33. 33.
    Barritt J, Willadsen S, Brenner C, Cohen J. Cytoplasmic transfer in assisted reproduction. Hum Reprod Update. 2001;7:428–35.CrossRefPubMedGoogle Scholar
  34. 34.
    Athayde Wirka K, Chen AA, Conaghan J, Ivani K, Gvakharia M, Behr B, et al. A typical embryo phenotypes identified by time-lapse microscopy: high prevalence and association with embryo development. Fertil Steril. 2014;101:1637–48.CrossRefPubMedGoogle Scholar
  35. 35.
    Yang ST, Shi JX, Gong F, Zhang SP, Lu CF, Tan K, et al. Cleavage pattern predicts developmental potential of day 3 human embryos produced by IVF. Reprod Biomed Online. 2015;30:625–34.CrossRefPubMedGoogle Scholar
  36. 36.
    Fragouli E, Wells D. Aneuploidy in the human blastocyst. Cytogenet Genome Res. 2011;133:149–59.CrossRefPubMedGoogle Scholar
  37. 37.
    Capalbo A, Rienzi L, Cimadomo D, Maggiulli R, Elliott T, Wright G, et al. Correlation between standard blastocyst morphology, euploidy and implantation: an observational study in two centers involving 956 screened blastocysts. Hum Reprod. 2014;29:1173–81.CrossRefPubMedGoogle Scholar
  38. 38.
    Howe K, FitzHarris G. Recent insights into spindle function in mammalian oocytes and early embryos. Biol Reprod. 2013;89:71.CrossRefPubMedGoogle Scholar
  39. 39.
    Jones KT, Lane SI. Molecular causes of aneuploidy in mammalian eggs. Development. 2013;140:3719–30.CrossRefPubMedGoogle Scholar
  40. 40.
    Jones KT. Meiosis in oocytes: predisposition to aneuploidy and its increased incidence with age. Hum Reprod Update. 2008;14:143–58.CrossRefPubMedGoogle Scholar
  41. 41.
    Zhang J, Zhuang G, Zeng Y, Grifo J, Acosta C, Shu Y, et al. Pregnancy derived from human zygote pronuclear transfer in a patient who had arrested embryos after IVF. Reprod Biomed Online. 2016;33:529–33.CrossRefPubMedGoogle Scholar
  42. 42.
    Zhang J. Revisiting germinal vesicle transfer as a treatment for aneuploidy in infertile women with diminished ovarian reserve. J Assist Reprod Genet. 2015;32:313–7.CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2017

Authors and Affiliations

  • Shuo-Ping Zhang
    • 1
    • 2
  • Chang-Fu Lu
    • 1
    • 2
    • 3
  • Fei Gong
    • 1
    • 2
    • 3
  • Ping-Yuan Xie
    • 1
    • 4
  • Liang Hu
    • 1
    • 2
    • 3
    • 4
  • Shun-Ji Zhang
    • 2
  • Guang-Xiu Lu
    • 1
    • 2
    • 3
    • 4
  • Ge Lin
    • 1
    • 2
    • 3
    • 4
  1. 1.Institute of Reproductive and Stem Cell EngineeringCentral South UniversityChangshaPeople’s Republic of China
  2. 2.Reproductive and Genetic Hospital of CITIC-XiangyaChangshaChina
  3. 3.Key laboratory of Reproductive and Stem Cell EngineeringMinistry of HealthChangshaChina
  4. 4.National Engineering and Research Center of Human Stem CellChangshaChina

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