Blastocyst utilization rates after continuous culture in two commercial single-step media: a prospective randomized study with sibling oocytes
- 341 Downloads
The aim of this study is to determine whether blastocyst utilization rates are different after continuous culture in two different commercial single-step media.
This is a paired randomized controlled trial with sibling oocytes conducted in infertility patients, aged ≤40 years with ≥10 oocytes retrieved assigned to blastocyst culture and transfer. Retrieved oocytes were randomly allocated to continuous culture in either Sage one-step medium (Origio) or Continuous Single Culture (CSC) medium (Irvine Scientific) without medium renewal up to day 5 post oocyte retrieval. Main outcome measure was the proportion of embryos suitable for clinical use (utilization rate).
A total of 502 oocytes from 33 women were randomly allocated to continuous culture in either Sage one-step medium (n = 250) or CSC medium (n = 252). Fertilization was performed by either in vitro fertilization or intracytoplasmic sperm injection, and embryo transfers were performed on day 5. Two patients had all blastocysts frozen due to the occurrence of severe ovarian hyperstimulation syndrome. Fertilization and cleavage rates, as well as embryo quality on day 3, were similar in the two media. Blastocyst utilization rates (%, 95% CI) [55.4% (46.4–64.1) vs 54.7% (44.9–64.6), p = 0.717], blastocyst formation rates [53.6% (44.6–62.5) vs 51.9 (42.2–61.6), p = 0.755], and proportion of good quality blastocysts [36.8% (28.1–45.4) vs 36.1% (27.2–45.0), p = 0.850] were similar in Sage one-step and CSC media, respectively.
Continuous culture of embryos in Sage one-step and CSC media is associated with similar blastocyst development and utilization rates. Both single-step media appear to provide adequate support during in vitro preimplantation embryo development. Whether these observations are also valid for other continuous single medium protocols remains to be determined.
Clinical trial registration number: NCT02302638.
KeywordsEmbryo culture Single step Continuous culture Blastocyst formation Utilization rate
The authors wish to thank Mrs. Maria Banti for embryology work and Mrs. G. Stavropoulou and Mrs. Ioanna Voulgaris for patient coordination.
IAS conceived the study, performed acquisition, analysis and interpretation of data, writing and revision of the manuscript, and embryology work. EMK, GTL, and CAV participated in the analysis and interpretation of data and writing and revision of the manuscript. GKP and BCT participated in the interpretation of data and revision of the manuscript. TGL had the general supervision of the study, participated in study design, analysis and interpretation of data, writing and revision of the manuscript, and performed clinical work. All authors read and approved the final manuscript.
Compliance with ethical standards
Sage one-step medium was provided by Origio, and CSC medium was provided by Biocare Europe.
Conflict of interest
The authors declare that they have no conflict of interest.
All procedures performed were in accordance with the ethical standards of the institutional research committee and with the 1964 Helsinki Declaration and its later amendments or comparable ethical standards.
Informed consent was obtained from all individual participants included in the study.
- 3.Machtinger R, Racowsky C. Culture systems: single step. In: Smith GD, Swain JE, Pool TB, editors. Embryo culture. Methods in molecular biology, 912. Humana Press; 2012. p. 199–209.Google Scholar
- 11.Lainas TG, Sfontouris IA, Zorzovilis IZ, Petsas GK, Lainas GT, Alexopoulou E, et al. Flexible GnRH antagonist protocol versus GnRH agonist long protocol in patients with polycystic ovary syndrome treated for IVF: a prospective randomised controlled trial (RCT). Hum Reprod. 2010;25:683–9.CrossRefPubMedGoogle Scholar
- 12.Mannaerts B, Van Kuijk J, Griesinger G. Prediction of OHSS in patients treated with corifollitropin alfa or rFSH in a GnRH antagonist protocol. Hum Reprod. 2012;27:ii26–i8.Google Scholar
- 14.Kolibianakis EM, Papanikolaou EG, Tournaye H, Camus M, Van Steirteghem AC, Devroey P. Triggering final oocyte maturation using different doses of human chorionic gonadotropin: a randomized pilot study in patients with polycystic ovary syndrome treated with gonadotropin-releasing hormone antagonists and recombinant follicle-stimulating hormone. Fertil Steril. 2007;88:1382–8.CrossRefPubMedGoogle Scholar
- 16.Hong KH, Lee H, Forman EJ, Upham KM, Scott RT Jr. Examining the temperature of embryo culture in in vitro fertilization: a randomized controlled trial comparing traditional core temperature (37°C) to a more physiologic, cooler temperature (36°C). Fertil Steril. 2014;102:767–73.CrossRefPubMedGoogle Scholar
- 19.Gardner D, Schoolcraft W. In vitro culture of the human blastocyst. In: Jansen R, Mortimer D, editors. Towards Reproductive Certainty: Infertility and Genetics beyond 1999. Carnforth: Parthenon publishing; 1999. p. 378–88.Google Scholar
- 25.Sfontouris IA, Kolibianakis EM, Lainas GT, Zorzovilis IZ, Petsas GK, Lainas TG. Blastocyst development in single-step versus sequential culture media: a prospective randomized study with sibling oocytes. Hum Reprod. 2014;29:i2.Google Scholar
- 28.Paternot G, Debrock S, D’Hooghe TM, Spiessens C. Early embryo development in a sequential versus single medium: a randomized study. Reprod Biol Endocrinol. 2010;8.Google Scholar
- 29.Sfontouris IA, Kolibianakis EM, Lainas GT, Petsas GK, Tarlatzis BC, Lainas TG. Blastocyst development in a single medium compared to sequential media. Reprod Sci. 2017:1933719116687653.Google Scholar
- 30.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;33:1261–72.CrossRefPubMedPubMedCentralGoogle Scholar
- 36.Bontekoe S, Heineman MJ, Johnson N, Blake D. Adherence compounds in embryo transfer media for assisted reproductive technologies. Cochrane Database Syst Rev. 2014;2:Cd007421.Google Scholar
- 41.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;30:1577–81.CrossRefPubMedPubMedCentralGoogle Scholar
- 42.Cohen J, Gilligan A, Esposito W, Schimmel T, Dale B. Ambient air and its potential effects on conception in vitro. Hum Reprod. 1997;12 Suppl 8:1742–9.Google Scholar
- 43.Hall J, Gilligan A, Schimmel T, Cecchi M, Cohen J. The origin, effects and control of air pollution in laboratories used for human embryo culture. Hum Reprod. 1998;13 Suppl 4:146–55.Google Scholar
- 44.Merton JS, Vermeulen ZL, Otter T, Mullaart E, de Ruigh L, Hasler JF. Carbon-activated gas filtration during in vitro culture increased pregnancy rate following transfer of in vitro-produced bovine embryos. Theriogenology. 2007;67 Suppl 7:1233–8.Google Scholar
- 45.Kleijkers SH, van Montfoort AP, Bekers O, Coonen E, Derhaag JG, Evers JL, et al. Ammonium accumulation in commercially available embryo culture media and protein supplements during storage at 2-8 °C and during incubation at 37oC. Hum Reprod. 2016;31:1192–9.Google Scholar