Early embryo development in a sequential versus single medium: a randomized study

Goedele Paternot, Sophie Debrock, Thomas M D'Hooghe, Carl Spiessens, Goedele Paternot, Sophie Debrock, Thomas M D'Hooghe, Carl Spiessens

Abstract

Background: The success of in vitro fertilization techniques is defined by multiple factors including embryo culture conditions, related to the composition of the culture medium. In view of the lack of solid scientific data and in view of the current general belief that sequential media are superior to single media, the aim of this randomized study was to compare the embryo quality in two types of culture media.

Methods: In this study, the embryo quality on day 3 was measured as primary outcome. In total, 147 patients younger than 36 years treated with IVF/ICSI during the first or second cycle were included in this study. Embryos were randomly cultured in a sequential (group A) or a single medium (group B) to compare the embryo quality on day 1, day 2 and day 3. The embryo quality was compared in both groups using a Chi-square test with a significance level of 0.05.

Results: At day 1, the percentage of embryos with a cytoplasmic halo was higher in group B (46%) than in group A (32%). At day 2, number of blastomeres, degree of fragmentation and the percentage of unequally sized blastomeres were higher in group B than in group A. At day 3, a higher percentage of embryos had a higher number of blastomeres and unequally sized blastomeres in group B. The number of good quality embryos (GQE) was comparable in both groups. The embryo utilization rate was higher in group B (56%) compared to group A (49%).

Conclusions: Although, no significant difference in the number of GQE was found in both media, the utilization rate was significantly higher when the embryos were cultured in the single medium compared to the sequential medium. The results of this study have a possible positive effect on the cumulative cryo-augmented pregnancy rate.

Trial registration number: NCT01094314.

References

    1. Aoki VW, Wilcox AL, Peterson CM, Parker-Jones K, Hatasaka HH, Gibson M, Huang I, Carell DT. Comparison of four media types during 3-day human IVF embryo culture. Reprod BioMed Online. 2005;10:600–606. doi: 10.1016/S1472-6483(10)61667-7.
    1. Hentemann M, Bertheussen K. New media for culture of blastocyst. Fertil Steril. 2009;91:878–883. doi: 10.1016/j.fertnstert.2007.12.011.
    1. Summers MC, Biggers JD. Chemically defined media and the culture of mammalian preimplantation embryos: historical perspective and current issues. Hum Reprod. 2003;9:557–582. doi: 10.1093/humupd/dmg039.
    1. Gardner DK, Lane M. Allevation of the '2-cell block' and development to the blastocyst of CFI mouse embryos: role of amino acids, EDTA and physical parameters. Hum Reprod. 1996;11:2703–2711.
    1. Gardner DK, Lane M. Culture of viable human blastocysts in defined sequential serum-free media. Hum Reprod. 1998;13:148–159. doi: 10.1093/humrep/13.12.3434.
    1. Gardner DK, Lane M. Towards a single embryo transfer. Reprod BioMed Online. 2003;6:470–481. doi: 10.1016/S1472-6483(10)62170-0.
    1. Lane M, Gardner DK. Embryo culture medium: which is the best? Best Res Clin Obstet Gyn. 2007;21:83–100. doi: 10.1016/j.bpobgyn.2006.09.009.
    1. Biggers JD, Summers MC. Choosing a culture medium: making informed choices. Fertil Steril. 2008;90:473–483. doi: 10.1016/j.fertnstert.2008.08.010.
    1. Gardner DK. Culture and selection of viable human blastocysts: a feasible proposition of human IVF. Hum Reprod update. 1997;3:367–382. doi: 10.1093/humupd/3.4.367.
    1. Pool TB. An update on embryo culture for human assisted reproductive technology: media, performance, and safety. Semin Reprod Med. 2002;23:309–318. doi: 10.1055/s-2005-923388.
    1. Gardner DK, Lane M. Amino acids and ammonium production regulate mouse embryo development in culture. Biol Reprod. 1993;48:377–385. doi: 10.1095/biolreprod48.2.377.
    1. Lane M, Gardner DK. Increase in postimplantation development of cultured mouse embryos by amino acids and induction of fetal retardation and encephaly by ammonium ions. J Reprod Fertil. 1994;102:305–312.
    1. Gardner DK, Pool TB, Lane M. Embryo nutrition and energy metabolism and its relationship to embryo growth, differentiation, and viability. Semin Reprod Med. 2000;18:205–218. doi: 10.1055/s-2000-12559.
    1. Biggers J, Racowsky C. The development of fertilized human ova to the blastocyst stage in KSOM AA medium: is a two-step protocol necessary? RBM Online. 2002;5:133–140.
    1. Xella S, Marsella T, Tagliasacchi D, Giulini S, La Marca A, Tirelli A, Volpe A. Embryo quality and implantation rate in two different culture media: ISM1 versus Universal IVF Medium. Fertil Steril. 2010;93(6):1859–63. doi: 10.1016/j.fertnstert.2008.12.030.
    1. Macklon NS, Pieters MHEC, Hassan MA, Jeucken PHM, Eijkemans MJC, Fauser BCJM. A prospective randomized comparison of sequential versus monoculture systems for in-vitro human blastocyst development. Hum Reprod. 2002;17:2700–2705. doi: 10.1093/humrep/17.10.2700.
    1. Sepulveda S, Garcia J, Arriagi E, Diaz J, Noriega-Portella L, Noriega-Hoces L. In vitro development and pregnancy outcome for human embryos cultured in either a single medium or in a sequential media system. Fertil Steril. 2009;91:1765–1770. doi: 10.1016/j.fertnstert.2008.02.169.
    1. Reed ML, Hamic A, Thompson DJ, Caperton CL. Continuous uninterrupted single medium culture without medium renewal versus sequential media culture: a sibling embryo study. Fertil Steril. 2009;92:1783–1789. doi: 10.1016/j.fertnstert.2009.05.008.
    1. Debrock S, Melotte C, Spiessens C, Peeraer K, Vanneste E, Meeuwis L, Meuleman C, Frijns JP, Vermeersch JR, D'Hooghe TM. Pre-implantation genetic screening for aneuploidy of embryos after in vitro fertilization in women aged at least 35 years: a prospective randomized trial. Fertil Steril. 2010;93:364–373. doi: 10.1016/j.fertnstert.2008.10.072.
    1. Gardner DK, Schoolcraft WB. In: vitro culture of the human blastocyst. Towards Reproductive Certainty: infertility and Genetics beyond 1999. Jansen R, Mortimer D, editor. Carnforth, UK: Parthenon Publishing; 1999. pp. 378–388.
    1. Zegers-Hochschild F, Adamson GD, de Mouzon J, Ishihara O, Mansour R, Nygren K, Sullivan E, van der Poel S. The international committee for monitoring assisted reproductive technology (ICMART) and the world health organisation (WHO) revised glossary on ART technology. Hum Reprod. 2009;24:2683–2687. doi: 10.1093/humrep/dep343.
    1. Bavister BD, Squirrell JM. Mitochondrial distribution and function in oocytes and early embryos. Hum Reprod. 2000;15:189–198. doi: 10.1093/humrep/15.1.189.
    1. Salumets A, Hydén-Granskog C, Suikkari AM, Tiitinen A, Tuuri T. The predictive value of pronuclear morphology of zygotes in the assessment of human embryo quality. Hum Reprod. 2001;16:2177–2181. doi: 10.1093/humrep/16.10.2177.
    1. Ebner T, Moser M, Sommergruber M, Gaiswinkler U, Wiesinger R, Puchner M, Tews G. Presence, but not type or degree of extension, of a cytoplasmic halo has a significant influence on preimplantation development and implantation behaviour. Hum Reprod. 2003;18:2406–2412. doi: 10.1093/humrep/deg452.
    1. Salumets A, Hydén-Granskog C, Mäkinen S, Suikkari AM, Tiitinen A, Tuuri T. Early cleavage predicts the viability of human embryos in elective single embryo transfer procedures. Hum Reprod. 2003;18:821–825. doi: 10.1093/humrep/deg184.
    1. Van Montfoort APA, Dumoulin JCM, Kester ADM, Evers JLH. Early cleavage is a valuable addition to existing embryo selection parameters: a study using single embryo transfers. Hum Reprod. 2004;19:2103–2108. doi: 10.1093/humrep/deh385.
    1. Ben-Yosef D, Amit A, Schwartz T, Cohen T, Mei-Raz N, Carmon A, Lessing JB, Yaron Y. Prospective randomized comparison of two embryo culture systems: P1 medium by Irvine Scientific and the COOK IVF medium. J Assist Reprod Genet. 2004;21:291–295. doi: 10.1023/B:JARG.0000043702.35570.56.

Source: PubMed

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