Does culture condition of reduced oxygen pressure helps in embryo quality? A prospective randomized study


  • Deepak Patil Department of Obstetrics and Gynaecology, Command Hospital Air Force, Bangalore, Karnataka, India



Embryo culture, Fragmentation rate, Vitrification


Background: Embryo quality is of paramount importance in an in vitro fertilization and embryo transfer (IVF-ET) cycle. A healthy embryo leads to a robust pregnancy. Incubation in an ideal atmosphere is essential requirement in an IVF-ET cycle.

Methods: Traditionally carbon dioxide incubator with sensors for carbon dioxide levels and temperature has been used widely. We compared the quality of these embryos to incubation in hypoxic condition by addition of nitrogen. Oxygen levels were brought to 5% as compared to 21% in room air.

Results: Ladies with high body mass index (BMI) and advanced age had more embryos available for transfer. More embryos reached day 3 and day 5, thus increasing availability for cryopreservation. The fragmentation rate and fertilization failure were less. Pregnancy rate was definitely improved as compared to traditional incubation.

Conclusions: Hypoxic incubation condition leads to better embryo health. This translates into improved and efficient cycle outcome.


Dow K. The men who made the breakthrough: How the British press represented Patrick Steptoe and Robert Edwards in 1978. Reprod Biomed Soc Online. 2017;4:59-67.

Fischer B, Bavister BD. Oxygen tension in the oviduct and uterus of rhesus monkeys, hamsters and rabbits. J Reprod Fertil. 1993;99(2):673-9.

Noda Y, Goto Y, Umaoka Y, Shiotani M, Nakayama T, Mori T. Culture of human embryos in alpha modification of Eagle's medium under low oxygen tension and low illumination. Fertil Steril. 1994;62(5):1022-7.

García-Martínez S, Sánchez Hurtado MA, Gutiérrez H, Sánchez Margallo FM, Romar R, Latorre R, Coy P, López Albors O. Mimicking physiological O2 tension in the female reproductive tract improves assisted reproduction outcomes in pig. Mol Hum Reprod. 2018;24(5):260-70.

Truong TT, Gardner DK. Antioxidants increase blastocyst cryosurvival and viability post-vitrification. Hum Reprod. 2020;35(1):12-23.

Tamura H, Jozaki M, Tanabe M, Shirafuta Y, Mihara Y, Shinagawa M, et al. Importance of Melatonin in Assisted Reproductive Technology and Ovarian Aging. Int J Mol Sci. 2020;21(3):1135.

Podkalicka P, Stępniewski J, Mucha O, Trojanowska NK, Dulak J, Łoboda A. Biomolecules. Hypoxia as a Driving Force of Pluripotent Stem Cell Reprogramming and Differentiation to Endothelial Cells. 2020;10(12):1614.

Bedaiwy MA, Falcone T, Mohamed MS, Aleem AA, Sharma RK, Worley SE, Thornton J, Agarwal A. Differential growth of human embryos in vitro: role of reactive oxygen species. Fertil Steril. 2004;82(3):593-600.

Cimadomo D, Fabozzi G, Vaiarelli A, Ubaldi N, Ubaldi FM, Rienzi L. Impact of Maternal Age on Oocyte and Embryo Competence. Front Endocrinol (Lausanne). 2018;9:327.

Bontekoe S, Mantikou E, van Wely M, Seshadri S, Repping S, Mastenbroek S. Low oxygen concentrations for embryo culture in assisted reproductive technologies. Cochrane Database Syst Rev. 2012;(7):CD008950.

Calzi F. Exposure of embryos to oxygen at low concentration in a cleavage stage transfer program: reproductive outcomes in a time-series analysis. Clin Lab. 2012;58(9-10):997-1003.

Dumoulin JC, Meijers CJ, Bras M, Coonen E, Geraedts JP, Evers JL. Effect of oxygen concentration on human in-vitro fertilization and embryo culture. Hum Reprod. 1999;14(2):465-9.

Sobrinho DBG. IVF/ICSI outcomes after culture of human embryos at low oxygen tension: a meta-analysis. Reprod Biol Endocrinol. 2011;9:143.

Waldenström U, Engström AB, Hellberg D, Nilsson S. Low-oxygen compared with high-oxygen atmosphere in blastocyst culture, a prospective randomized study. Fertil Steril. 2009;91(6):2461-5.

Montfoort APAV.Reduced oxygen concentration during human IVF culture improves embryo utilization and cumulative pregnancy rates per cycle. Hum Reprod Open. 2020;2020(1):036.

Belli M, Zhang L, Liu X, Donjacour A, Ruggeri E, Palmerini MG, Nottola SA, Macchiarelli G, Rinaudo P. Oxygen concentration alters mitochondrial structure and function in in vitro fertilized preimplantation mouse embryos. Hum Reprod. 2019;34(4):601-11.

Zhang XY, Xiong YM, Tan YJ, Wang L, Li R, Zhang Y, et al. Melatonin rescues impaired penetration ability of human spermatozoa induced by mitochondrial dysfunction. Reproduction. 2019;158(5):465-75.

Abad MR. The influence of oxygen concentration during embryo culture on obstetric and neonatal outcomes: a secondary analysis of a randomized controlled trial. Hum Reprod. 2020;35(9):2017-25.

Ruíz M. Effect of Group Embryo Culture under Low-Oxygen Tension in Benchtop Incubators on Human Embryo Culture: Prospective, Randomized, Controlled Trial. Reprod Sci. 2020;27(7):1522-33.

Bavister B. Oxygen concentration and preimplantation development. Reprod Biomed. 2004;9(5):484-6.

Guo N, Li Y, Ai J, Gu L, Chen W, Liu Q. Two different concentrations of oxygen for culturing precompaction stage embryos on human embryo development competence: a prospective randomized sibling-oocyte study. Int J Clin Exp Pathol. 2014;7(9):6191-8.

Plessis SSD, Hagenaar K, Lampiao F. The in vitro effects of melatonin on human sperm function and its scavenging activities on NO and ROS. Andrologia. 2010;42(2):112-6.






Original Research Articles