Skip to main content
SpringerLink
Log in

Unbalanced human embryos secrete more hyperglycosylated human chorionic gonadotrophin (hCG-H) than balanced ones

  • Embryo Biology
  • Published:
Journal of Assisted Reproduction and Genetics Aims and scope Submit manuscript

Abstract

Purpose

The aim of this study was to compare the levels of hyperglycosylated human chorionic gonadotropin (hCG-H) secreted from balanced and unbalanced human embryos.

Methods

Single-step culture media samples from 155 good quality embryos, derived from 90 good prognosis patients undergoing intracytoplasmic sperm injection (ICSI), were collected on the fifth day of embryo cultivation. All embryos were tested by next-generation sequencing (NGS) technique. The hCG-H levels in the culture media were evaluated by ELISA kit (Cusabio Biotech, CBS-E15803h) according to the manufacturer’s instructions. Statistical analysis was performed using SPSS v.21 (IBM Corp., Armonk, NY, USA).

Results

The NGS analysis revealed that 36% of the embryos (n = 56) were balanced, and 64% of the embryos were unbalanced (n = 99). The presence of hCG-H was confirmed in all embryo culture media samples but was absent in the negative control. In addition, hCG-H concentration was significantly higher in the culture media from unbalanced embryos compared with the balanced ones (0.72 ± 0.30 mIU/ml vs. 0.62 ± 0.12 mIU/ml, p = 0.02, respectively). Furthermore, the mean levels of hCG-H were significantly increased in the samples from embryos with multiple abnormalities. Finally, the highest levels of hCG-H were expressed from embryos with monosomy of chromosome 11 (1.28 ± 0.04 mIU/ml) and those with trisomies of chromosomes 21 (2.23 mIU/ml) and 4 (1.02 ± 0.35 mIU/ml).

Conclusion

Our results suggest that chromosomal aberrations in human embryos are associated with an increased secretion of hCG-H. However, hCG-H concentration in embryo culture media as a single biomarker is not sufficient for an accurate selection of balanced embryos.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

References

  1. Chavez SL, Loewke KE, Han J, Moussavi F, Colls P, Munne S, et al. Dynamic blastomere behaviour reflects human embryo ploidy by the four-cell stage. Nat Commun. 2012;3:1251.

    Article  PubMed  Google Scholar 

  2. Findikli N, Kahraman S, Saglam Y, Beyazyurek C, Sertyel S, Karlikaya G, et al. Embryo aneuploidy screening for repeated implantation failure and unexplained recurrent miscarriage. Reprod BioMed Online. 2006;13(1):38–46.

    Article  CAS  PubMed  Google Scholar 

  3. Wang Y, Chen Y, Tian F, Zhang J, Song Z, Wu Y, et al. Maternal mosaicism is a significant contributor to discordant sex chromosomal aneuploidies associated with noninvasive prenatal testing. Clin Chem. 2014;60(1):251–9.

    Article  PubMed  Google Scholar 

  4. Campbell A, Fishel S, Bowman N, Duffy S, Sedler M, Hickman CF. Modelling a risk classification of aneuploidy in human embryos using non-invasive morphokinetics. Reprod BioMed Online. 2013;26(5):477–85.

    Article  PubMed  Google Scholar 

  5. Hassold T, Hunt P. To err (meiotically) is human: the genesis of human aneuploidy. Nat Rev Genet. 2001;2(4):280–91.

    Article  CAS  PubMed  Google Scholar 

  6. Liu L, Li Y, Li S, Hu N, He Y, Pong R, et al. Comparison of next-generation sequencing systems. J Biomed Biotechnol. 2012;2012:251364.

    PubMed  PubMed Central  Google Scholar 

  7. Cimadomo D, Capalbo A, Ubaldi FM, Scarica C, Palagiano A, Canipari R, et al. The impact of biopsy on human embryo developmental potential during preimplantation genetic diagnosis. Biomed Res Int. 2016;2016:7193075.

    Article  PubMed  PubMed Central  Google Scholar 

  8. Botros L, Sakkas D, Seli E. Metabolomics and its application for non-invasive embryo assessment in IVF. Mol Hum Reprod. 2008;14(12):679–90.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Liang B, Gao Y, Xu J, Song Y, Xuan L, Shi T, et al. Raman profiling of embryo culture medium to identify aneuploid and euploid embryos. Fertil Steril. 2019;111(4):753–62 e1.

    Article  PubMed  Google Scholar 

  10. Butler SA, Luttoo J, Freire MO, Abban TK, Borrelli PT, Iles RK. Human chorionic gonadotropin (hCG) in the secretome of cultured embryos: hyperglycosylated hCG and hCG-free beta subunit are potential markers for infertility management and treatment. Reprod Sci. 2013;20(9):1038–45.

    Article  PubMed  Google Scholar 

  11. Cole LA. New discoveries on the biology and detection of human chorionic gonadotropin. Reprod Biol Endocrinol. 2009;7:8.

    Article  PubMed  PubMed Central  Google Scholar 

  12. Kovalevskaya G, Kakuma T, Schlatterer J, O'Connor JF. Hyperglycosylated HCG expression in pregnancy: cellular origin and clinical applications. Mol Cell Endocrinol. 2007;260-262:237–43.

    Article  CAS  PubMed  Google Scholar 

  13. Mock P, Kovalevskaya G, O'Connor JF, Campana A. Choriocarcinoma-like human chorionic gonadotrophin (HCG) and HCG bioactivity during the first trimester of pregnancy. Hum Reprod. 2000;15(10):2209–14.

    Article  CAS  PubMed  Google Scholar 

  14. Cole LA. Hyperglycosylated hCG. Placenta. 2007;28(10):977–86.

    Article  CAS  PubMed  Google Scholar 

  15. Guibourdenche J, Handschuh K, Tsatsaris V, Gerbaud P, Leguy MC, Muller F, et al. Hyperglycosylated hCG is a marker of early human trophoblast invasion. J Clin Endocrinol Metab. 2010;95(10):E240–4.

    Article  CAS  PubMed  Google Scholar 

  16. Kajihara T, Uchino S, Suzuki M, Itakura A, Brosens JJ, Ishihara O. Human chorionic gonadotropin confers resistance to oxidative stress-induced apoptosis in decidualizing human endometrial stromal cells. Fertil Steril. 2011;95(4):1302–7.

    Article  CAS  PubMed  Google Scholar 

  17. Fogle RH, Li A, Paulson RJ. Modulation of HOXA10 and other markers of endometrial receptivity by age and human chorionic gonadotropin in an endometrial explant model. Fertil Steril. 2010;93(4):1255–9.

    Article  CAS  PubMed  Google Scholar 

  18. Yang H, Lei CX, Zhang W. Human chorionic gonadotropin (hCG) regulation of galectin-3 expression in endometrial epithelial cells and endometrial stromal cells. Acta Histochem. 2013;115(1):3–7.

    Article  CAS  PubMed  Google Scholar 

  19. O'Connor JF, Ellish N, Kakuma T, Schlatterer J, Kovalevskaya G. Differential urinary gonadotrophin profiles in early pregnancy and early pregnancy loss. Prenat Diagn. 1998;18(12):1232–40.

    Article  CAS  PubMed  Google Scholar 

  20. Kovalevskaya G, Birken S, Kakuma T, Ozaki N, Sauer M, Lindheim S, et al. Differential expression of human chorionic gonadotropin (hCG) glycosylation isoforms in failing and continuing pregnancies: preliminary characterization of the hyperglycosylated hCG epitope. J Endocrinol. 2002;172(3):497–506.

    Article  CAS  PubMed  Google Scholar 

  21. Sutton-Riley JM, Khanlian SA, Byrn FW, Cole LA. A single serum test for measuring early pregnancy outcome with high predictive value. Clin Biochem. 2006;39(7):682–7.

    Article  CAS  PubMed  Google Scholar 

  22. Sasaki Y, Ladner DG, Cole LA. Hyperglycosylated human chorionic gonadotropin and the source of pregnancy failures. Fertil Steril. 2008;89(6):1781–6.

    Article  CAS  PubMed  Google Scholar 

  23. Grossman K, Cordina M, Ross JA. Is there a role for hyperglycosylated hCG in the diagnosis of ectopic pregnancy in women with a pregnancy of unknown location? J Reprod Med. 2017;62(1–2):15–20.

    CAS  PubMed  Google Scholar 

  24. Bahado-Singh RO, Oz AU, Kingston JM, Shahabi S, Hsu CD, Cole L. The role of hyperglycosylated hCG in trophoblast invasion and the prediction of subsequent pre-eclampsia. Prenat Diagn. 2002;22(6):478–81.

    Article  CAS  PubMed  Google Scholar 

  25. Keikkala E, Koskinen S, Vuorela P, Laivuori H, Romppanen J, Heinonen S, et al. First trimester serum placental growth factor and hyperglycosylated human chorionic gonadotropin are associated with pre-eclampsia: a case control study. BMC Pregnancy Childbirth. 2016;16(1):378.

    Article  PubMed  PubMed Central  Google Scholar 

  26. Cole LA. Proportion hyperglycosylated hCG: a new test for discriminating gestational trophoblastic diseases. Int J Gynecol Cancer. 2014;24(9):1709–14.

    Article  PubMed  Google Scholar 

  27. Ramu S, Acacio B, Adamowicz M, Parrett S, Jeyendran RS. Human chorionic gonadotropin from day 2 spent embryo culture media and its relationship to embryo development. Fertil Steril. 2011;96(3):615–7.

    Article  CAS  PubMed  Google Scholar 

  28. Xiao-Yan C, Jie L, Dang J, Tao L, Xin-Ru L, Guang-Lun Z. A highly sensitive electrochemiluminescence immunoassay for detecting human embryonic human chorionic gonadotropin in spent embryo culture media during IVF-ET cycle. J Assist Reprod Genet. 2013;30(3):377–82.

    Article  PubMed  Google Scholar 

  29. Wang H, Zhang R, Han D, Liu C, Cai J, Bi Y, et al. Association of human chorionic gonadotropin level in embryo culture media with early embryo development. Nan Fang Yi Ke Da Xue Xue Bao. 2014;34(7):1039–41 1047.

    PubMed  Google Scholar 

  30. Parvanov D, Nikolova D, Ganeva R, Nikolova K, Vasileva M, Rangelov I, et al. Maternal age and blastocyst quality do not influence the embryo production of hyperglycosylated human chorionic gonadotropin. Fertil Steril. 2019;112(3):e130.

    Article  Google Scholar 

  31. Stamenov G, Nikolova K, Vasileva M, Rangelov I, Ganeva R, Pancheva M, et al. Hyperglycosylated human chorionic gonadotropin in blastocyst culture medium of male and female human embryos. Fertil Steril. 2019;112(3):e126.

    Article  Google Scholar 

  32. McCallie BR, Parks JC, Denomme Tignanelli M, Schoolcraft WB, Katz-Jaffe M. RNA sequencing of aneuploid blastocysts revealed enriched pathways associated with compromised implantation potential. Fertil Steril. 2016;106(3):е23.

    Article  Google Scholar 

  33. Spencer K. Evaluation of an assay of the free beta-subunit of choriogonadotropin and its potential value in screening for Down's syndrome. Clin Chem. 1991;37:809–14.

    Article  CAS  PubMed  Google Scholar 

  34. Spencer K, Macri JN, Aitken DA, Connor JM. Free beta-hCG as first-trimester marker for fetal trisomy. Lancet. 1992;339:1480.

    Article  CAS  PubMed  Google Scholar 

  35. Bogart MH, Pandian MR, Jones OW. Abnormal maternal serum chorionic gonadotropin levels in pregnancies with fetal chromosome abnormalities. Prenat Diagn. 1987;7:623–30.

    Article  CAS  PubMed  Google Scholar 

  36. Bindra R, Liao VHA, Spencer K, Nicolaides KH. One stop clinic for assessment of risk for trisomy 21 at 11–14 weeks: a prospective study of 15030 pregnancies. Ultrasound Obstet Gynecol. 2002;20:219–25.

    Article  CAS  PubMed  Google Scholar 

  37. Bernd E. Glaubitz Ralf. First-trimester screening: an overview. J Histochem Cytochem. 2005;53(3):281–3.

    Article  Google Scholar 

  38. Cole LA, Dai D, Butler SA, Leslie KK, Kohorn EI. Gestational trophoblastic diseases: 1. Pathophysiology of hyperglycosylated hCG. Gynecol Oncol. 2006;102:144–9.

    Google Scholar 

  39. Cole LA, Khanlian SA, Riley JM, Butler SA. Hyperglycosylated hCG in gestational implantation and in choriocarcinoma and testicular germ cell malignancy tumorigenesis. J Reprod Med. 2006;51:919–29.

    CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Nadezhda Women’s Health Hospital, 3 “Blaga vest” Street, Sofia, Bulgaria

    Dimitar Parvanov, Rumiana Ganeva, Kristina Nikolova, Magdalena Vasileva, Ivaylo Rangelov, Maria Pancheva, Maria Serafimova & Georgi Stamenov

  2. Department of Medical Genetics, Medical Faculty, Medical University – Sofia, Sofia, Bulgaria

    Dragomira Nikolova, Rada Staneva & Savina Hadjidekova

  3. Centre for Endocrinology and Reproductive Medicine, Rome, Italy

    Fabio Scarpellini

Authors
  1. Dimitar Parvanov
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Dragomira Nikolova
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Rumiana Ganeva
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Kristina Nikolova
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Magdalena Vasileva
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Ivaylo Rangelov
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Maria Pancheva
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Maria Serafimova
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Rada Staneva
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Savina Hadjidekova
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Fabio Scarpellini
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Georgi Stamenov
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

D.P., D.N., and G.S. conceived the experiment; D.P., R.G., K.N., M.V, I.R., M.P., M.S., R.S., and S.H. conducted the experiment; D.P., D.N., R.G., R.S., F.S., and S.H. analyzed the results. D.P., D.N., and R.G. wrote the main manuscript text and prepared tables and figures. R.S., S.H. D.P, R.G., and G.S. edited the manuscript and made its final revision. All authors critically reviewed and approved the final version of the manuscript.

Corresponding author

Correspondence to Dimitar Parvanov.

Ethics declarations

Conflict of interest

The authors declare that they have no conflicts of interest.

Ethical approval

The protocol of the study and the informed consent forms have been approved by the IRB committee of the Nadezhda University Hospital and conforms to the ethical principles of the Declaration of Helsinki for medical research involving human subjects.

Informed consent

All experiments were non-invasive and the women have given a written informed consent for participation in the research.

Additional information

Publisher’s note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Parvanov, D., Nikolova, D., Ganeva, R. et al. Unbalanced human embryos secrete more hyperglycosylated human chorionic gonadotrophin (hCG-H) than balanced ones. J Assist Reprod Genet 37, 1341–1348 (2020). https://doi.org/10.1007/s10815-020-01776-9

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10815-020-01776-9

Keywords

Search

Navigation