Skip to main content
SpringerLink
Log in

Embryo Culture Medium Has No Impact on Mosaicism Rates: a Sibling Oocyte Study

  • Embryology: Original Article
  • Published:
Reproductive Sciences Aims and scope Submit manuscript

Abstract

Human embryos cultured in vitro can contain two or more cytogenetically distinct cell lineages known as “chromosomal mosaicism”. Since mosaicism is produced by mitotic errors after fertilization occurs, culture conditions might contribute to mosaicism origins. Many studies demonstrated that euploidy rates are not affected by culture media; however, whether oocytes cultured under continuous culture media (CCM) or sequential culture media (SCM) has a higher risk of mosaicism occurring remains unsolved. Therefore, this study aims to determine whether mosaicism rates differ when sibling oocytes are cultured in CCM or SCM. A single center observational study was performed including 6072 sibling oocytes. Mature oocytes (MII) were inseminated and cultured in CCM (n = 3,194) or SCM (n = 2,359) until blastocyst stage for trophectoderm (TE) biopsy on day (D) 5, D6, or D7 for preimplantation genetic testing analysis with a semi-automated next-generation sequencing. Mosaicism was classified as low (30–50%) or high (50–80%) based on the percentage of abnormal cells constitution detected in TE samples. As a result, 426 women with a mean age of 34.7 ± 6.4 years were included in the study. Fertilization rates were comparable between CCM and SCM (74.0% vs 72.0%, p = 0.091). Although total blastulation rate and usable blastocyst rate (biopsied blastocysts) were significantly higher in CCM than SCM (75.3 % vs. 70.3%, p < 0.001 and 58.0% vs. 54.5%, p = 0.026), euploidy rates did not differ significantly (45.2% vs. 45.7%, p = 0.810, respectively). Mosaicism rate was not significantly different for blastocysts cultured in CCM or SCM (4.7% vs. 5.1%, p = 0.650), neither the proportion of low or high mosaic rates (3.7% vs. 4.4%, p = 0.353 and 1.0% vs. 0.7%, p = 0.355, respectively). Hence, it was concluded that CCM or SCM does not have an impact on mosaicism rate of embryos cultured until the blastocyst stage.

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

Similar content being viewed by others

Data Availability

Data are available as per request.

Code Availability

None.

References

  1. Viotti M. Preimplantation genetic testing for chromosomal abnormalities: aneuploidy, mosaicism, and structural rearrangements. Genes (Basel). 2020;11(6):602. https://doi.org/10.3390/genes11060602.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Popovic M, Dhaenens L, Boel A, Menten B, Heindryckx B. Chromosomal mosaicism in human blastocysts: the ultimate diagnostic dilemma. Hum Reprod Update. 2020;26(3):313–34. https://doi.org/10.1093/humupd/dmz050.

    Article  CAS  PubMed  Google Scholar 

  3. Capalbo A, Hoffmann ER, Cimadomo D, Ubaldi FM, Rienzi L. Human female meiosis revised: new insights into the mechanisms of chromosome segregation and aneuploidies from advanced genomics and time-lapse imaging. Hum Reprod Update. 2017;23:706–22. https://doi.org/10.1093/humupd/dmx026.

    Article  CAS  PubMed  Google Scholar 

  4. Munné S, Wells D. Detection of mosaicism at blastocyst stage with the use of high-resolution next-generation sequencing. Fertil Steril. 2017;107:1085–91. https://doi.org/10.1016/j.fertnstert.2017.03.024.

    Article  PubMed  Google Scholar 

  5. Yang M, Rito T, Metzger J, Naftaly J, Soman R, Hu J, Albertini DF, Barad DH, Brivanlou AH, Gleicher N. Depletion of aneuploid cells in human embryos and gastruloids. Nat Cell Biol. 2021;23(4):314–21. https://doi.org/10.1038/s41556-021-00660-7.

    Article  CAS  PubMed  Google Scholar 

  6. Goodrich D, Tao X, Bohrer C, Lonczak A, Xing T, Zimmerman R, Zhan Y, Scott RT Jr, Treff NR. A randomized and blinded comparison of qPCR and NGS-based detection of aneuploidy in a cell line mixture model of blastocyst biopsy mosaicism. J Assist Reprod Genet. 2016;33(11):1473–80. https://doi.org/10.1007/s10815-016-0784-3.

    Article  PubMed  PubMed Central  Google Scholar 

  7. Coll L, Parriego M, Mateo S, García-Monclús S, Rodríguez I, Boada M, Coroleu B, Polyzos NP, Vidal F, Veiga A. Prevalence, types and possible factors influencing mosaicism in IVF blastocysts: results from a single setting. Reprod Biomed Online. 2021;42(1):55–65. https://doi.org/10.1016/j.rbmo.2020.09.025.

    Article  CAS  PubMed  Google Scholar 

  8. Armstrong A, Miller J, Quinn M, Nguyen AV, Kwan L, Kroener L. To mask or not to mask mosaicism? The impact of reporting embryo mosaicism on reproductive potential. J Assist Reprod Genet. 2022;39(9):2035–42. https://doi.org/10.1007/s10815-022-02576-z.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Ata B, Popovic M, Fatemi H. Correct assessment and interpretation of results determines the accuracy of any diagnostic test, and PGT-A is no exception. Hum Reprod. 2022;37(9):2214–6. https://doi.org/10.1093/humrep/deac150.

    Article  PubMed  Google Scholar 

  10. Abhari S, Kawwass JF. Pregnancy and Neonatal Outcomes after Transfer of Mosaic Embryos: A Review. J Clin Med. 2021;10(7):1369. https://doi.org/10.3390/jcm10071369.

    Article  PubMed  PubMed Central  Google Scholar 

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

    Article  CAS  PubMed  Google Scholar 

  12. Morbeck DE, Baumann NA, Oglesbee D. Composition of single-step media used for human embryo culture. Fertil Steril. 2017;107(4):1055–1060.e1. https://doi.org/10.1016/j.fertnstert.2017.01.007.

    Article  PubMed  Google Scholar 

  13. Quinn P. Culture systems: sequential. Methods Mol Biol. 2012;912:211–30. https://doi.org/10.1007/978-1-61779-971-6_13.

    Article  CAS  PubMed  Google Scholar 

  14. Werner MD, Hong KH, Franasiak JM, Forman EJ, Reda CV, Molinaro TA, Upham KM, Scott RT Jr. 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 

  15. Abdala A, Elkhatib I, Bayram A, Arnanz A, El-Damen A, Melado L, Lawrenz B, Coughlan C, Garrido N, Fatemi HM, De Munck N. 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. 2021;38(8):2199–207. https://doi.org/10.1007/s10815-021-02187-0.

    Article  PubMed  PubMed Central  Google Scholar 

  16. Quinn MM, Marsh P, Ribeiro S, Simbulan RK, Hickman C, Berntsen J, Rosen MP. Aneuploidy rates and morphokinetic parameters of embryos cultured in distinct culture media: a sibling oocyte study. Hum Reprod. 2022;37(2):226–34. https://doi.org/10.1093/humrep/deab253.

    Article  PubMed  Google Scholar 

  17. 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 

  18. Cimadomo D, Scarica C, Maggiulli R, Orlando G, Soscia D, Albricci L, Romano S, Sanges F, Ubaldi FM, Rienzi L. 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 

  19. Lin PY, Lee CI, Cheng EH, Huang CC, Lee TH, Shih HH, Pai YP, Chen YC, Lee MS. Clinical outcomes of single mosaic embryo transfer: high-level or low-level mosaic embryo, does it matter? J Clin Med. 2020;9(6):1695. https://doi.org/10.3390/jcm9061695.

    Article  PubMed  PubMed Central  Google Scholar 

  20. Greco E, Minasi MG, Fiorentino F. Healthy babies after intrauterine transfer of mosaic aneuploid blastocysts. N. Engl. J. Med. 2015;373:2089–90.

    Article  PubMed  Google Scholar 

  21. Capalbo A, Poli M, Rienzi L, Girardi L, Patassini C, Fabiani M, Cimadomo D, Benini F, Farcomeni A, Cuzzi J, Rubio C, Albani E, Sacchi L, Vaiarelli A, Figliuzzi M, Findikli N, Coban O, Boynukalin FK, Vogel I, et al. Mosaic human preimplantation embryos and their developmental potential in a prospective, non-selection clinical trial. Am J Hum Genet. 2021;108(12):2238–47. https://doi.org/10.1016/j.ajhg.2021.11.002.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Zhang L, Wei D, Zhu Y, Gao Y, Yan J, Chen ZJ. Rates of live birth after mosaic embryo transfer compared with euploid embryo transfer. J Assist Reprod Genet. 2019;36(1):165–72. https://doi.org/10.1007/s10815-018-1322-2.

    Article  PubMed  Google Scholar 

  23. De Munck N, El Khatib I, Abdala A, El-Damen A, Bayram A, Arnanz A, Melado L, Lawrenz B, Fatemi HM. Intracytoplasmic sperm injection is not superior to conventional IVF in couples with non-male factor infertility and preimplantation genetic testing for aneuploidies (PGT-A). Hum Reprod. 2020;35(2):317–27. https://doi.org/10.1093/humrep/deaa002.

    Article  CAS  PubMed  Google Scholar 

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

    Google Scholar 

  25. 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 

  26. García-Pascual CM, Navarro-Sánchez L, Navarro R, Martínez L, Jiménez J, Rodrigo L, Simón C, Rubio C. Optimized NGS approach for detection of aneuploidies and mosaicism in PGT-A and imbalances in PGT-SR. Genes (Basel). 2020;11(7):724. https://doi.org/10.3390/genes11070724.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Sfontouris IA, Martins WP, Nastri CO, Viana IG, Navarro PA, Raine-Fenning N, van der Poel S, Rienzi L, Racowsky C. 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 

  28. Swain JE. Can culture media impact preimplantation embryo aneuploidy? Fertil Reprod. 2021;3(04):120–4. https://doi.org/10.1142/S2661318221500183.

    Article  Google Scholar 

  29. 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 

  30. Munné S, Kaplan B, Frattarelli JL, Child T, Nakhuda G, Shamma FN, Silverberg K, Kalista T, Handyside AH, Katz-Jaffe M, Wells D, Gordon T, Stock-Myer S. Willman S; STAR Study Group. Preimplantation genetic testing for aneuploidy versus morphology as selection criteria for single frozen-thawed embryo transfer in good-prognosis patients: a multicenter randomized clinical trial. Fertil Steril. 2019;112(6):1071–1079.e7. https://doi.org/10.1016/j.fertnstert.2019.07.1346.

    Article  CAS  PubMed  Google Scholar 

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

    Article  PubMed  PubMed Central  Google Scholar 

  32. 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;28:634–41. https://doi.org/10.1093/humrep/des462.

    Article  CAS  PubMed  Google Scholar 

  33. Hardarson T, Bungum M, Conaghan J, Meintjes M, Chantilis SJ, Molnar L, Gunnarsson K, Wikland M. Noninferiority, randomized, controlled trial comparing embryo development using media developed for sequential or undisturbed culture in a time-lapse setup. Fertil Steril. 2015;104(6):1452–9.e1-4. https://doi.org/10.1016/j.fertnstert.2015.08.037.

    Article  PubMed  Google Scholar 

  34. Fragouli E, Alfarawati S, Simkins M, Cutts G, Spath K, Babariya D, Kubikova N, Rubistello L, Munne S, Wells D. Factors affecting embryonic mosaicism. Hum Rep. 2017;32:i49.

    Google Scholar 

  35. Hozyen M, Elshimy A, Mohamed N, Abdeltawab A, Elkhader H, Zaki H. Sequential versus single step culture media in relation to euploidy rates in pgtai program: preliminary data. Fertil Steril. 2020;114(3):e146–7.

    Article  Google Scholar 

  36. Behr B, Lathi R, Cinnioglu C, Kort J. The effect of changing culture media on aneuploidy rates seen in preimplantation embryos. Hum. Rep. 2018;33:i241.

    Google Scholar 

  37. Nakhuda G, Jing C, Butler R, Guimond C, Hitkari J, Taylor E, Tallon N, Yuzpe A. Frequencies of chromosome-specific mosaicisms in trophoectoderm biopsies detected by next-generation sequencing. Fertil Steril. 2018;109(5):857–65. https://doi.org/10.1016/j.fertnstert.2018.01.011.

    Article  CAS  PubMed  Google Scholar 

  38. Spinella F, Fiorentino F, Biricik A, Bono S, Ruberti A, Cotroneo E, Baldi M, Cursio E, Minasi MG, Greco E. Extent of chromosomal mosaicism influences the clinical outcome of in vitro fertilization treatments. Fertil Steril. 2018;109(1):77–83. https://doi.org/10.1016/j.fertnstert.2017.09.025.

    Article  PubMed  Google Scholar 

  39. Fragouli E, Alfarawati S, Spath K, Babariya D, Tarozzi N, Borini A, Wells D. Analysis of implantation and ongoing pregnancy rates following the transfer of mosaic diploid-aneuploid blastocysts. Hum Genet. 2017;136(7):805–19. https://doi.org/10.1007/s00439-017-1797-4.

    Article  CAS  PubMed  Google Scholar 

  40. Practice Committee and Genetic Counseling Professional Group (GCPG) of the American Society for Reproductive Medicine. Clinical management of mosaic results from preimplantation genetic testing for aneuploidy (PGT-A) of blastocysts: a committee opinion. Fertil Steril. 2020;114(2):246–54. https://doi.org/10.1016/j.fertnstert.2020.05.014.

    Article  CAS  Google Scholar 

  41. Ai X, Shi Y, Liu LW, Xu Y, Zhang H, Liu Y, Wang J, Ding C, Cai B, Zhou C, Xu Y. Risk factors related to chromosomal mosaicism in human blastocysts. Reprod Biomed Online. 2022;45(1):54–62. https://doi.org/10.1016/j.rbmo.2022.02.016.

    Article  CAS  PubMed  Google Scholar 

  42. Lee CI, Cheng EH, Lee MS, Lin PY, Chen YC, Chen CH, Huang LS, Huang CC, Lee TH. Healthy live births from transfer of low-mosaicism embryos after preimplantation genetic testing for aneuploidy. J Assist Reprod Genet. 2020;37(9):2305–13. https://doi.org/10.1007/s10815-020-01876-6.

    Article  PubMed  PubMed Central  Google Scholar 

  43. ESHRE Working Group on Chromosomal Mosaicism, De Rycke M, Capalbo A, Coonen E, Coticchio G, Fiorentino F, Goossens V, Mcheik S, Rubio C, Sermon K, Sfontouris I, Spits C, Vermeesch JR, Vermeulen N, Wells D, Zambelli F, Kakourou G. ESHRE survey results and good practice recommendations on managing chromosomal mosaicism. Hum Reprod Open. 2022;2022(4):hoac044. https://doi.org/10.1093/hropen/hoac044.

    Article  PubMed Central  Google Scholar 

  44. Munné S, Spinella F, Grifo J, Zhang J, Beltran MP, Fragouli E, Fiorentino F. Clinical outcomes after the transfer of blastocysts characterized as mosaic by high resolution Next Generation Sequencing- further insights. Eur J Med Genet. 2020 63(2):103741. https://doi.org/10.1016/j.ejmg.2019.103741.

    Article  PubMed  Google Scholar 

  45. Lawrenz B, El Khatib I, Liñán A, Bayram A, Arnanz A, Chopra R, De Munck N, Fatemi HM. The clinicians´ dilemma with mosaicism-an insight from inner cell mass biopsies. Hum Reprod. 2019;34(6):998–1010. https://doi.org/10.1093/humrep/dez055.

    Article  CAS  PubMed  Google Scholar 

  46. Victor AR, Griffin DK, Brake AJ, Tyndall JC, Murphy AE, Lepkowsky LT, Lal A, Zouves CG, Barnes FL, McCoy RC, Viotti M. Assessment of aneuploidy concordance between clinical trophectoderm biopsy and blastocyst. Hum Reprod. 2019;34(1):181–92. https://doi.org/10.1093/humrep/dey327.

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We thank our biostatistician Dr. Rachna Patel for the statistical analysis and critical review of the manuscript.

Author information

Authors and Affiliations

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

    Andrea Abdala, Ibrahim Elkhatib, Aşina Bayram, Ahmed El-Damen, Laura Melado, Barbara Lawrenz, Human M. Fatemi & Daniela Nogueira

  2. Obstetrical Department, Women’s University Hospital Tuebingen, Tuebingen, Germany

    Barbara Lawrenz

  3. Inovie Fertilité, Toulouse, France

    Daniela Nogueira

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. Ahmed El-Damen
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Laura Melado
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Barbara Lawrenz
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Human M. Fatemi
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Daniela Nogueira
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

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

Corresponding author

Correspondence to Andrea Abdala.

Ethics declarations

Ethics Approval

This study was approved by the local ethical committee (REFA070a).

Consent to Participate

No consent was needed. Personal identification was coded to preserve patients’ identity.

Consent to Publish

Data can be published.

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

Rights and permissions

Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

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. Embryo Culture Medium Has No Impact on Mosaicism Rates: a Sibling Oocyte Study. Reprod. Sci. 30, 3296–3304 (2023). https://doi.org/10.1007/s43032-023-01276-y

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s43032-023-01276-y

Keywords

Search

Navigation