Reproductive Medicine and Biology

, Volume 9, Issue 3, pp 163–168 | Cite as

A study of the effect of an extremely low oxygen concentration on the development of human embryos in assisted reproductive technology

  • Koji NakagawaEmail author
  • Asako Shirai
  • Yayoi Nishi
  • Rie Sugiyama
  • Yasushi Kuribayashi
  • Rikikazu Sugiyama
  • Masato Inoue
Original Article



To determine whether embryos cultured with a low oxygen level (2%) brought about beneficial effects on the outcome of ART.


This is a sequential case–control embryo-culture study. Embryos were cultured either with a gas mixture containing 2% O2, 5% CO2, and 93% N2 (low-oxygen group) or 5% O2, 5% CO2, and 90% N2 (conventional group). From January 2008 to September 2008, 873 fertilized oocytes were obtained from 250 patients in the low-oxygen group and from October 2008 to March 2009, 730 fertilized oocytes were obtained from 213 patients in the conventional group. The outcomes of ART were compared between two groups.


The cleavage rate in the low-oxygen group (94.4%) was similar to that (94.7%) in the conventional group. The mean number of blastomeres on Day 3 in the low-oxygen group (mean ± SE) was 6.5 ± 1.9, and this was significantly lower than in the conventional group (6.8 ± 1.9, p < 0.05). Moreover, the low-oxygen group produced worse quality embryos, on the basis of the significantly higher embryo grade 2.1 ± 0.6 versus 1.9 ± 0.6, p < 0.001, in 5% oxygen. The pregnancy and miscarriage rates in the low-oxygen group were 22.3 and 20.8%, respectively, which were statistically similar to the outcomes in the conventional group.


Overall, culture of embryos at the low oxygen level did not significantly improve ART results compared with embryos grown in 5% oxygen. The study suggests that a low oxygen level worsens embryo morphology but does not impair embryo viability.


ART Embryo development Embryo quality High oxygen Low oxygen 



We wish to thank Dr Alex Lopata for his helpful advice and for editing the paper.


  1. 1.
    Dumoulin JC, Vanvuchelen RC, Land JA, Pieters MH, Geraedts JP, Evers JL. Effect of oxygen concentration on in vitro fertilization and embryo culture in the human and the mouse. Fertil Steril. 1995;63:115–9.PubMedGoogle Scholar
  2. 2.
    Dumoulin JC, Meijers CJ, Bras M, Coonen E, Geraedts J, Evers JL. Effect of oxygen concentration on human in vitro fertilization and embryo transfer. Hum Reprod. 1999;14:465–9.CrossRefPubMedGoogle Scholar
  3. 3.
    Bavister BD. Culture of preimplantation embryos: facts and artifacts. Hum Reprod Update. 1995;1:91–148.CrossRefPubMedGoogle Scholar
  4. 4.
    Fischer B, Bavister BD. Oxygen tension in the oviduct and uterus of rhesus monkeys, hamsters and rabbits. J Reprod Fertil. 1993;99:673–9.CrossRefPubMedGoogle Scholar
  5. 5.
    Goto Y, Noda Y, Mori T, Nakano M. Increased generation of reactive oxygen species in embryos cultured in vitro. Free Radic Biol Med. 1993;15:69–75.CrossRefPubMedGoogle Scholar
  6. 6.
    Sugiyama R, Nakagawa K, Nishi Y, Ezaki K, Sugiyama R, Inoue M. The dilemma faced by patients who undergo single embryo transfer. Reprod Biol Med. 2009;8:33–7.CrossRefGoogle Scholar
  7. 7.
    Nakagawa K, Ohgi S, Kojima R, Sugawara K, Horikawa T, Ito M, et al. Recombinant-FSH has more effective potent than urinary human menopausal gonadotropin in ovarian hyperstimulation for ART treatment. Reprod Biol Med. 2007;6:27–32.CrossRefGoogle Scholar
  8. 8.
    Nakagawa K, Ohgi S, Kojima R, Ito M, Horikawa T, Irahara M, et al. Reduction of perifollicular arterial blood flow resistance after hCG administration is a good indicator of the recovery of mature oocytes in ART treatment. J Assist Reprod Genet. 2006;23:433–8.CrossRefPubMedGoogle Scholar
  9. 9.
    Nakagawa K, Yamano S, Moride N, Yamashita M, Yoshizawa M, Aono T. Effect of activation with Ca ionophore A23187 and puromycin on the development of human oocytes that failed to fertilize after intracytoplasmic sperm injection. Fertil Steril. 2001;76:148–52.CrossRefPubMedGoogle Scholar
  10. 10.
    Veeck LL. Atlas of the human oocyte and early conceptus, vol 2. Baltimore: Williams & Wilkins Co; 1991.Google Scholar
  11. 11.
    Steptoe PC, Edward RG. Birth after the reimplantation of a human embryo. Lancet. 1978;2:366.CrossRefPubMedGoogle Scholar
  12. 12.
    Kwon HC, Yang HW, Hwang KJ, Yoo HJ, Kim MS, Lee CH, et al. Effects of low oxygen condition on the generation of reactive oxygen species and the development in mouse embryos cultured in vitro. J Obstet Gynecol Res. 1999;25:359–66.CrossRefGoogle Scholar
  13. 13.
    Lim JM, Reggio BC, Godke KA, Hansel W. Development of in-vitro derived bovine embryos cultured in 5% CO2 in air or 5% O2, 5% CO2 and 90% N2. Hum Reprod. 1999;14:458–64.CrossRefPubMedGoogle Scholar
  14. 14.
    Batt PA, Gardner DK, Cameron WN. Oxygen concentration and protein source affect the development of preimplantation goat embryos in vitro. Reprod Fertil Dev. 1991;3:601–7.CrossRefPubMedGoogle Scholar
  15. 15.
    Quinn P, Harlow G. The effect of oxygen on the development of preimplantation mouse embryo in vitro. J Exp Zool. 1978;206:73–80.CrossRefPubMedGoogle Scholar
  16. 16.
    Nagao Y, Iijima R, Saeki K. Interaction between embryos and culture conditions during in vitro development of bovine early embryos. Zygote. 2008;16:127–33.CrossRefPubMedGoogle Scholar
  17. 17.
    Berthelot F, Terqui M. Effects of oxygen, CO2/pH and medium in the in vitro development of individually cultured procine one- and two-cell embryos. Reprod Nutr Dev. 1996;36:241–51.CrossRefPubMedGoogle Scholar
  18. 18.
    Ciray HN, Aksoy T, Yaramanci K, Karayaka I, Bahceci M. In vitro culture under physiologic oxygen concentration improves blastocyst yield and quality: a prospective randomized survey on sibling oocytes. Fertil Steril. 2009;91(4 suppl):1459–61.CrossRefPubMedGoogle Scholar
  19. 19.
    Kea B, Gebhardt J, Watt J, Westphal LM, Lathi RB, Milki AA, et al. Effect of reduced oxygen concentrations on the outcome of in vitro fertilization. Fertil Steril. 2007;87:213–6.CrossRefPubMedGoogle Scholar
  20. 20.
    Waldenstrom U, Engstrom AB, Hellberg D, Nilsson S. Low-oxygen compared with high-oxygen atmosphere in blastocyst culture, a prospective randomized study. Fertil Steril. 2008 (Epub ahead of print).Google Scholar
  21. 21.
    Thompson JGE, Simpson AC, Pugh PA, Donnelly PE, Tervit HR. Effect of oxygen concentration on in vitro development of preimplantation sheep and cattle embryos. J Reprod Fertil. 1990;89:573–8.CrossRefPubMedGoogle Scholar

Copyright information

© Japan Society for Reproductive Medicine 2010

Authors and Affiliations

  • Koji Nakagawa
    • 1
    Email author
  • Asako Shirai
    • 1
  • Yayoi Nishi
    • 1
  • Rie Sugiyama
    • 1
  • Yasushi Kuribayashi
    • 1
  • Rikikazu Sugiyama
    • 1
  • Masato Inoue
    • 1
  1. 1.Division of Reproductive MedicineSugiyama ClinicTokyoJapan

Personalised recommendations