Skip to main content
SpringerLink
Log in

Euploidy rates are not affected when embryos are cultured in a continuous (CCM) or sequential culture medium (SCM): a sibling oocyte study

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

Abstract

Purpose

To determine if euploidy rates and embryo development differ when blastocysts are cultured in CCM or SCM.

Method

A single-center retrospective observational study was performed from September 2018 to March 2019. Patients [23–46 years] with at least four fresh mature oocytes (MII) without severe male factor infertility were included. Sibling MII were injected and cultured in Global®Total®LP (CCM) or Sage Quinn’s Advantage® Cleavage and Blastocyst media (SCM) under 6% CO2, 5% O2, and 89% N2. Fertilization, cleavage, day (D) 5 blastulation, usable blastocyst (blastocysts biopsied/normally fertilized oocytes), and euploidy rates were recorded. Blastocysts were graded prior to trophectoderm (TE) biopsy on D5, 6, or 7 for genetic testing and mitochondrial DNA (mtDNA) quantification.

Results

According to clinical practice, 1452 MII were randomly distributed: 751 in CCM and 701 in SCM. No differences were observed in fertilization and cleavages rates for CCM and SCM (77.4% vs 75.5%, p = 0.429 and 97.6% vs 99.1%, p = 0.094, respectively). Blastulation rate on D5 was higher in CCM (70.6% vs 62.2, p = 0.009); however, usable blastocyst rates were comparable (CCM: 58.3% vs SCM: 56.7%, p = 0.625). From a Poisson regression model adjusted for confounding factors, euploidy rates were not different between media (aOR = 1.18, [0.94–1.48], p = 0.157). Euploid blastocyst’s mtDNA values were similar (CCM: 32.2, [30.5, 34.1] and SCM: 33.5, [31.8, 35.2], p = 0.345) and top-quality blastocysts (AA/BA) were increased in SCM (OR=1.04, [1.00–1.09], p = 0.037).

Conclusion

Under controlled in vitro conditions, euploidy rates and embryo development are comparable when embryos are cultured in CCM or SCM.

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

Access this article

Log in via an institution

Subscribe and save

Springer+ Basic
EUR 32.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or Ebook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

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

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2

Similar content being viewed by others

Data availability

Available per request.

References

  1. Castillo CM, Harper J, Roberts SA, O’Neill HC, Johnstone ED, Brison DR. The impact of selected embryo culture conditions on ART treatment cycle outcomes: a UK national study. Hum Reprod Open. 2020;2020(1):hoz031. https://doi.org/10.1093/hropen/hoz031.

    Article  PubMed  PubMed Central  Google Scholar 

  2. Sunde A, Brison D, Dumoulin J, Harper J, Lundin K, Magli MC, et al. Time to take human embryo culture seriously. Hum Reprod. 2016;31(10):2174–82. https://doi.org/10.1093/humrep/dew157.

    Article  PubMed  Google Scholar 

  3. Ménézo Y, Lichtblau I, Elder K. New insights into human pre-implantation metabolism in vivo and in vitro. J Assist Reprod Genet. 2013;30(3):293–303. https://doi.org/10.1007/s10815-013-9953-9.

    Article  PubMed  PubMed Central  Google Scholar 

  4. Gardner DK, Lane M, Calderon I, Leeton J. Environment of the preimplantation human embryo in vivo: metabolite analysis of oviduct and uterine fluids and metabolism of cumulus cells. Fertil Steril. 1996;65(2):349–53. https://doi.org/10.1016/s0015-0282(16)58097-2.

    Article  CAS  PubMed  Google Scholar 

  5. Gardner DK. Changes in requirements and utilization of nutrients during mammalian preimplantation embryo development and their significance in embryo culture. Theriogenology. 1998;49(1):83–102. https://doi.org/10.1016/s0093-691x(97)00404-4.

    Article  CAS  PubMed  Google Scholar 

  6. Summers MC, Biggers JD. Chemically defined media and the culture of mammalian preimplantation embryos: historical perspective and current issues. Hum Reprod Update. 2003;9(6):557–82. https://doi.org/10.1093/humupd/dmg039.

    Article  CAS  PubMed  Google Scholar 

  7. Costa-Borges N, Bellés M, Meseguer M, Galliano D, Ballesteros A, Calderón G. Blastocyst development in single medium with or without renewal on day 3: a prospective cohort study on sibling donor oocytes in a time-lapse incubator. Fertil Steril. 2016;105(3):707–13. https://doi.org/10.1016/j.fertnstert.2015.11.038.

    Article  PubMed  Google Scholar 

  8. Sepúlveda S, Garcia J, Arriaga E, Diaz J, Noriega-Portella L, Noriega-Hoces L. In vitro development and pregnancy outcomes for human embryos cultured in either a single medium or in a sequential media system. Fertil Steril. 2009;91(5):1765–70. https://doi.org/10.1016/j.fertnstert.2008.02.169.

    Article  PubMed  Google Scholar 

  9. Sfontouris IA, Martins WP, Nastri CO, Viana IG, Navarro PA, Raine-Fenning N, et al. Blastocyst culture using single versus sequential media in clinical IVF: a systematic review and meta-analysis of randomized controlled trials. J Assist Reprod Genet. 2016;33(10):1261–72. https://doi.org/10.1007/s10815-016-0774-5.

    Article  PubMed  PubMed Central  Google Scholar 

  10. Ciray HN, Aksoy T, Goktas C, Ozturk B, Bahceci M. Time-lapse evaluation of human embryo development in single versus sequential culture media--a sibling oocyte study. J Assist Reprod Genet. 2012;29(9):891–900. https://doi.org/10.1007/s10815-012-9818-7.

    Article  PubMed  PubMed Central  Google Scholar 

  11. López-Pelayo I, Gutiérrez-Romero JM, Armada AIM, Calero-Ruiz MM, Acevedo-Yagüe PJM, et al. JBRA Assist Reprod. 2018;22(2):128–33. https://doi.org/10.5935/1518-0557.20180024.

    Article  PubMed  PubMed Central  Google Scholar 

  12. VerMilyea M, Rios C, Abu Maizar AM, James RM, Picou A, Werland HJ, et al. Stress relief: can continuous culture in a low lactate culture media reduce numerical chromosomal abnormalities and therefore improve euploidy rates? Fertil Steril. 2018;110:e361–2. https://doi.org/10.10161/j.fertnstert.2018.07.1009.

    Article  Google Scholar 

  13. Deng J, Zhao Q, Cinnioglu C, Kayali R, Lathi RB, Behr B. The impact of culture conditions on blastocyst formation and aneuploidy rates: a comparison between single-step and sequential media in a large academic practice. J Assist Reprod Genet. 2020;37(1):161–9. https://doi.org/10.1007/s10815-019-01621-8.

    Article  PubMed  PubMed Central  Google Scholar 

  14. Desai N, Flyckt R, Austin CM. Can culture medium affect morpho kinetics, embryo dysmorphisms and euploidy rate? Fertil Steril. 2017;108(3):e160. https://doi.org/10.1016/j.fertnstert.2017.07.482.

    Article  Google Scholar 

  15. Werner MD, Hong KH, Franasiak JM, Forman EJ, Reda CV, Molinaro TA, et al. Sequential versus Monophasic Media Impact Trial (SuMMIT): a paired randomized controlled trial comparing a sequential media system to a monophasic medium. Fertil Steril. 2016;105(5):1215–21. https://doi.org/10.1016/j.fertnstert.2016.01.005.

    Article  CAS  PubMed  Google Scholar 

  16. Fabozzi G, Albricci L, Cimadomo D, Amendola MG, Sanges F, Maggiulli R, et al. Blastulation rates of sibling oocytes in two IVF cultur emedia: an evidence-based workflow to implement newly commercialized products. Reprod BioMed Online. 2020; Article in press. https://doi.org/10.1016/j.rbmo.2020.10.017.

  17. Fragouli E, Spath K, Alfarawati S, Kaper F, Craig A, Michel CE, et al. Altered levels of mitochondrial DNA are associated with female age, aneuploidy, and provide an independent measure of embryonic implantation potential. PLoS Genet. 2015;11(6):e1005241. https://doi.org/10.1371/journal.pgen.1005241.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Fragouli E, McCaffrey C, Ravichandran K, Spath K, Grifo JA, Munné S, et al. Clinical implications of mitochondrial DNA quantification on pregnancy outcomes: a blinded prospective non-selection study. Hum Reprod. 2017;32(11):2340–7. https://doi.org/10.1093/humrep/dex292 Erratum in: Hum Reprod. 2019 Apr 1;34(4):782.

    Article  CAS  PubMed  Google Scholar 

  19. Wells D. Mitochondrial DNA quantity as a biomarker for blastocyst implantation potential. Fertil Steril. 2017;108(5):742–7. https://doi.org/10.1016/j.fertnstert.2017.10.007.

    Article  CAS  PubMed  Google Scholar 

  20. Victor AR, Brake AJ, Tyndall JC, Griffin DK, Zouves CG, Barnes FL, et al. Accurate quantitation of mitochondrial DNA reveals uniform levels in human blastocysts irrespective of ploidy, age, or implantation potential. Fertil Steril. 2017;107(1):34–42.e3. https://doi.org/10.1016/j.fertnstert.2016.09.028.

    Article  CAS  PubMed  Google Scholar 

  21. Treff NR, Zhan Y, Tao X, Olcha M, Han M, Rajchel J, et al. Levels of trophectoderm mitochondrial DNA do not predict the reproductive potential of sibling embryos. Hum Reprod. 2017;32(4):954–62. https://doi.org/10.1093/humrep/dex034.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Sighinolfi G, Grisendi V, La Marca A. How to personalize ovarian stimulation in clinical practice. J Turk Ger Gynecol Assoc. 2017;18(3):148–53. https://doi.org/10.4274/jtgga.2017.0058.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Gardner DK, Schoolcraft WB. In vitro culture of human blastocyst. In: Jansen R, Mortimer D, editors. Towards Reproductive Certainty: Fertility and Genetics Beyond 1999. Cornforth: Parthenon Publishing; 1999. p. 378–88.

    Google Scholar 

  24. Thompson SM, Onwubalili N, Brown K, Jindal SK, McGovern PG. Blastocyst expansion score and trophectoderm morphology strongly predict successful clinical pregnancy and live birth following elective single embryo blastocyst transfer (eSET): a national study. J Assist Reprod Genet. 2013;30(12):1577–81. https://doi.org/10.1007/s10815-013-0100-4.

    Article  PubMed  PubMed Central  Google Scholar 

  25. Honnma H, Baba T, Sasaki M, Hashiba Y, Ohno H, Fukunaga T, et al. Trophectoderm morphology significantly affects the rates of ongoing pregnancy and miscarriage in frozen-thawed single-blastocyst transfer cycle in vitro fertilization. Fertil Steril. 2012;98(2):361–7. https://doi.org/10.1016/j.fertnstert.2012.05.014.

    Article  PubMed  Google Scholar 

  26. Aoyama N, Kato K. Trophectoderm biopsy for preimplantation genetic test and technical tips: A review. Reprod Med Biol. 2020;19(3):222–31. https://doi.org/10.1002/rmb2.12318.

    Article  PubMed  PubMed Central  Google Scholar 

  27. Phillips NR, Sprouse ML, Roby RK. Simultaneous quantification of mitochondrial DNA copy number and deletion ratio: a multiplex real-time PCR assay. Sci Rep. 2014;4:3887. https://doi.org/10.1038/srep03887.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Morbeck DE, Krisher RL, Herrick JR, Baumann NA, Matern D, Moyer T. Composition of commercial media used for human embryo culture. Fertil Steril. 2014;102(3):759–766.e9. https://doi.org/10.1016/j.fertnstert.2014.05.043.

    Article  CAS  PubMed  Google Scholar 

  29. Chronopoulou E, Harper JC. IVF culture media: past, present and future. Hum Reprod Update. 2015;21(1):39–55. https://doi.org/10.1093/humupd/dmu040.

    Article  CAS  PubMed  Google Scholar 

  30. Leese HJ, Whitear SL. Female tract environment and its relationship to ART media composition. In: Quinn P, editor. Culture media, solutions and systems in human ART. Cambridge: Cambridge University Press; 2014. p. 21–9. https://doi.org/10.1017/CBO9781139059053.004.

    Chapter  Google Scholar 

  31. Summers MC, Bird S, Mirzai FM, Thornhill A, Biggers JD. Human preimplantation embryo development in vitro: a morphological assessment of sibling zygotes cultured in a single medium or in sequential media. Hum Fertil (Camb). 2013;16(4):278–85. https://doi.org/10.3109/14647273.2013.806823.

    Article  Google Scholar 

  32. Swain JE. Optimizing the culture environment in the IVF laboratory: impact of pH and buffer capacity on gamete and embryo quality. Reprod BioMed Online. 2010;21(1):6–16. https://doi.org/10.1016/j.rbmo.2010.03.012.

    Article  PubMed  Google Scholar 

  33. Cimadomo D, Scarica C, Maggiulli R, Orlando G, Soscia D, Albricci L, et al. Continuous embryo culture elicits higher blastulation but similar cumulative delivery rates than sequential: a large prospective study. J Assist Reprod Genet. 2018;35(7):1329–38. https://doi.org/10.1007/s10815-018-1195-4.

    Article  PubMed  PubMed Central  Google Scholar 

  34. Biggers JD, McGinnis LK, Lawitts JA. One-step versus two-step culture of mouse preimplantation embryos: is there a difference? Hum Reprod. 2005;20(12):3376–84. https://doi.org/10.1093/humrep/dei228.

    Article  CAS  PubMed  Google Scholar 

  35. ESHRE Working group on Time-lapse technology, Apter S, Ebner T, Freour T, Guns Y, Kovacic B, et al. Good practice recommendations for the use of time-lapse technology†. Hum Reprod Open. 2020;2020(2):hoaa008. https://doi.org/10.1093/hropen/hoaa008.

    Article  PubMed Central  Google Scholar 

  36. Basile N, Morbeck D, García-Velasco J, Bronet F, Meseguer M. Type of culture media does not affect embryo kinetics: a time-lapse analysis of sibling oocytes. Hum Reprod. 2013;28(3):634–41. https://doi.org/10.1093/humrep/des462.

    Article  CAS  PubMed  Google Scholar 

  37. Schiewe MC, Whitney JB, Nugent N, Zozula S, Anderson RE. Human blastocyst development: A randomized comparison of sibling zygotes cultured in Vitrolife G-TLTM to Life Global® single-step media. J Pregnancy Reprod. 2018;2. https://doi.org/10.15761/JPR.1000144.

  38. Loutradis D, Drakakis P, Kallianidis K, Sofikitis N, Kallipolitis G, Milingos S, et al. Biological factors in culture media affecting in vitro fertilization, preimplantation embryo development, and implantation. Ann N Y Acad Sci. 2000;900:325–35. https://doi.org/10.1111/j.1749-6632.2000.tb06245.x.

    Article  CAS  PubMed  Google Scholar 

  39. Takashima A, Kimura F, Kishida K, Izuno M, Hirata K, et al. Comparison of embryo development and pregnancy rates in continuous single and sequential media cultures of sibling embryos. J Adv Med Sci Appl Technol. 2017;3(3):147–54. https://doi.org/10.32598/jamsat.3.3.147.

    Article  Google Scholar 

  40. Dieamant F, Petersen CG, Mauri AL, et al. Single versus sequential culture medium: which is better at improving ongoing pregnancy rates? A systematic review and meta-analysis. JBRA Assist Reprod. 2017;21(3):240–6. Published 2017 Sep 1. https://doi.org/10.5935/1518-0557.20170045.

    Article  PubMed  PubMed Central  Google Scholar 

  41. Franasiak JM, Forman EJ, Hong KH, Werner MD, Upham KM, Treff NR, et al. The nature of aneuploidy with increasing age of the female partner: a review of 15,169 consecutive trophectoderm biopsies evaluated with comprehensive chromosomal screening. Fertil Steril. 2014;101(3):656–663.e1. https://doi.org/10.1016/j.fertnstert.2013.11.004.

    Article  PubMed  Google Scholar 

  42. Swain JE, Carrell D, Cobo A, Meseguer M, Rubio C, Smith GD. Optimizing the culture environment and embryo manipulation to help maintain embryo developmental potential. Fertil Steril. 2016;105(3):571–87. https://doi.org/10.1016/j.fertnstert.2016.01.035 Erratum in: Fertil Steril. 2016 May;105(5):1377.

    Article  PubMed  Google Scholar 

  43. Diez-Juan A, Rubio C, Marin C, Martinez S, Al-Asmar N, Riboldi M, et al. Mitochondrial DNA content as a viability score in human euploid embryos: less is better. Fertil Steril. 2015;104(3):534–41.e1. https://doi.org/10.1016/j.fertnstert.2015.05.022.

    Article  CAS  PubMed  Google Scholar 

  44. Racowsky C, Vernon M, Mayer J, Ball GD, Behr B, Pomeroy KO, et al. Standardization of grading embryo morphology. J Assist Reprod Genet. 2010;27(8):437–9. https://doi.org/10.1007/s10815-010-9443-2.

    Article  PubMed  PubMed Central  Google Scholar 

  45. Gruber I, Klein M. Embryo culture media for human IVF: which possibilities exist? J Turk Ger Gynecol Assoc. 2011;12(2):110–7. https://doi.org/10.5152/jtgga.2011.25.

    Article  PubMed  PubMed Central  Google Scholar 

  46. Swain JE. Controversies in ART: can the IVF laboratory influence preimplantation embryo aneuploidy? Reprod BioMed Online. 2019;39(4):599–607. https://doi.org/10.1016/j.rbmo.2019.06.009.

    Article  CAS  PubMed  Google Scholar 

  47. Lane M, Gardner DK. Embryo culture medium: which is the best? Best Pract Res Clin Obstet Gynaecol. 2007;21(1):83–100. https://doi.org/10.1016/j.bpobgyn.2006.09.009.

    Article  PubMed  Google Scholar 

  48. Bouillon C, Leandri R, Desch L, Ernst A, Bruno C, Cerf C, et al. Does embryo culture medium influence the health and development of children born after in vitro fertilization? PLoS One. 2016;11(3):e0150857. https://doi.org/10.1371/journal.pone.0150857.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

We would like to thank Guillermo Moll�� Robles for his help and support with statistical evaluation and interpretation.

Author information

Authors and Affiliations

  1. ART Fertility Clinics, Abu Dhabi, United Arab Emirates

    Andrea Abdala, Ibrahim Elkhatib, Aşina Bayram, Ana Arnanz, Ahmed El-Damen, Laura Melado, Barbara Lawrenz, Carol Coughlan, Human M. Fatemi & Neelke De Munck

  2. IVI Foundation, Instituto de Investigación Sanitaria La Fe (IIS La Fe), Valencia, Spain

    Nicolas Garrido

  3. Obstetrical Department, Women’s university hospital Tuebingen, Tuebingen, Germany

    Barbara Lawrenz

Authors
  1. Andrea Abdala
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Ibrahim Elkhatib
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Aşina Bayram
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Ana Arnanz
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Ahmed El-Damen
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Laura Melado
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Barbara Lawrenz
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Carol Coughlan
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Nicolas Garrido
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Human M. Fatemi
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Neelke De Munck
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

ALA, ND, HF: conception and design of the study; ALA, ND, IE, AA, AD, AB, LM, BL, HF: oocyte retrieval; ND, IE, AA, AD, AB, ALA: IVF and ICSI procedure and embryo evaluations; ND, IE, AA, AD, AB, ALA: blastocyst biopsy, tubing, and freezing; ALA: construction of the database, NG: statistical analysis and data interpretation; ALA: drafting the manuscript; ND, IE, AA, AD, AB,CC, LM, BL, HF: critical review and final approval of the manuscript.

Corresponding author

Correspondence to Andrea Abdala.

Ethics declarations

Conflict of interest

The authors declare no competing interests.

Additional information

Publisher’s note

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

Supplementary information

ESM 1

(DOCX 16 kb).

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Abdala, A., Elkhatib, I., Bayram, A. et al. Euploidy rates are not affected when embryos are cultured in a continuous (CCM) or sequential culture medium (SCM): a sibling oocyte study. J Assist Reprod Genet 38, 2199–2207 (2021). https://doi.org/10.1007/s10815-021-02187-0

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10815-021-02187-0

Keywords

Search

Navigation