The study aims to summarize current knowledge on the use of oil in embryo culture systems, with a focus on proper management of different types of oil and possible impact on culture systems.
PubMed was used to search the MEDLINE database for peer-reviewed English-language original articles and reviews concerning the use of oil in embryo culture systems. Searches were performed by adopting “embryo,” “culture media,” “oil,” and “contaminants” as main terms. The most relevant publications were assessed and discussed critically.
Oils used in IVF are complex mixtures of straight-chain hydrocarbons, cyclic and aromatic hydrocarbons, and unsaturated hydrocarbons, whose precise composition influences their chemical and physical properties. Possible presence of contaminants suggests their storage at 4 °C in the dark to prevent peroxidation. Washing, generally performed by manufacturers prior to commercialization, may remove trace chemical contaminants. Oils reduce evaporation from culture media at rates depending on their chemical physical properties, culture system parameters, and incubator atmosphere. Contaminants — mainly metal ion and plastic components derived from refinement processes and storage — can pass to the aqueous phase of culture systems and affect embryo development.
Oils are essential components of culture systems. Their original quality and composition, storage, handling, and use can affect embryo development with significant efficiency and safety implications.
This is a preview of subscription content,to check access.
Access this article
Similar content being viewed by others
Gardner DK, Weissman A, Howles CM, Shoham Z (Eds). Textbook of Assisted Reproductive Techniques: Volume 2: Clinical Perspectives (5th ed.). CRC Press; 2018. https://doi.org/10.1201/9781351228244
Gardner DK, Kelley RL. Impact of the IVF laboratory environment on human preimplantation embryo phenotype. J Dev Origins Health Dis. 2017;8(4):418–35. https://doi.org/10.1017/S2040174417000368.
Brinster RL. A Method for in vitro cultivation of mouse ova from two-cell to blastocyst. Exp Cell Res. 1963;32(1):205–8. https://doi.org/10.1016/0014-4827(63)90093-4.
Ainsworth AJ, Fredrickson JR, Morbeck DE. Improved detection of mineral oil toxicity using an extended mouse embryo assay. J Assist Reprod Genet. 2017;34(3):391–7. https://doi.org/10.1007/s10815-016-0856-4.
Morbeck DE, Khan Z, Barnidge DR, Walker DL. Washing mineral oil reduces contaminants and embryotoxicity. Fertil Steril. 2010;94(7):2747–52. https://doi.org/10.1016/j.fertnstert.2010.03.067.
Provo MB, Herr C. Washed paraffin oil becomes toxic to mouse embryos upon exposure to sunlight. Theriogenology. 1998;49(1):214. https://doi.org/10.1016/S0093-691X(98)90567-2.
Otsuki J, Nagai Y, Chiba K. Peroxidation of mineral oil used in droplet culture is detrimental to fertilization and embryo development. Fertil Steril. 2007;88(3):741–3. https://doi.org/10.1016/j.fertnstert.2006.11.144.
EFSA Panel on contaminants in the food chian. Scientific Opinion on Mineral Oil Hydrocarbons in Food. Efsa J. 2012;10.
Morbeck DE, Leonard PH. Culture systems: mineral oil overlay. Methods Mol Biol. 2012;912:325–31. https://doi.org/10.1007/978-1-61779-971-6_18
Biedermann M, Fiselier K, Grob K. Aromatic hydrocarbons of mineral oil origin in foods: method for determining the total concentration and first results. J Agric Food Chem. 2009;57(19):8711–21. https://doi.org/10.1021/jf901375e
Pirow R, Blume A, Hellwig N, Herzler M, Huhse B, Hutzler C, Pfaff K, Thierse HJ, Tralau T, Vieth B, Luch A. Mineral oil in food, cosmetic products, and in products regulated by other legislations. Crit Rev Toxicol. 2019;49(9):742–89. https://doi.org/10.1080/10408444.2019.1694862
Fleming TP, Pratt HP, Braude PR. The use of mouse preimplantation embryos for quality control of culture reagents in human in vitro fertilization programs: a cautionary note. Fertil Steril. 1987;47(5):858–60. https://doi.org/10.1016/s0015-0282(16)59179-1
Lee S, Cho M, Kim E, Kim T, Lee C, Han J, et al. Renovation of a drop embryo cultures system by using refined mineral oil and the effect of glucose and/or hemoglobin added to a serum-free medium. J Vet Med Sci. 2004;66(1):63–6. https://doi.org/10.1292/jvms.66.63.
Swain JE, Schoolcraft WB, Bossert N, Batcheller AE. Media osmolality changes over 7 days following culture in a non-humidified benchtop incubator. Fertil Steril. 2016;106(3):e362. https://doi.org/10.1016/j.fertnstert.2016.07.1028.
Swain JE, Graham C, Kile R, Schoolcraft WB, Krisher RL. Media evaporation in a dry culture incubator; effect of dish, drop size and oil on media osmolality. Fertil Steril. 2018;110(4):e363–4. https://doi.org/10.1016/j.fertnstert.2018.07.1015.
Swain JE. Controversies in ART: considerations and risks for uninterrupted embryo culture. Reprod Biomed Online. 2019;39(1):19–26. https://doi.org/10.1016/j.rbmo.2019.02.009.
Del Gallego R, Albert C, Marcos J, Larreategui Z, Alegre L, Meseguer M. Humid vs. dry embryo culture conditions on embryo development: a continuous embryo monitoring assessment. Fertil Steril. 2018;110(4):e362–3. https://doi.org/10.1016/j.fertnstert.2018.07.1012.
Mestres E, García-Jiménez M, Casals A, Cohen J, Acacio M, Villamar A, et al. Factors of the human embryo culture system that may affect media evaporation and osmolality. Hum Reprod. 2021;36(3):605–13. https://doi.org/10.1093/humrep/deaa370.
Olds S, Stemm K, Wachter K, Wiemer K. Analysis of embryo culture media pH changes during incubator use and media evaporation under oil using a continuous pH monitoring system. Fertil Steril. 2015;104(3):e318–9. https://doi.org/10.1016/j.fertnstert.2015.07.997.
Yumoto K, Iwata K, Sugishima M, Yamauchi J, Nakaoka M, Matsumoto I, et al. Mineral oil viscosity affects the osmotic pressure of human embryonic culture medium microdrops in non-humidified incubators. Fertility and Sterility. 2018;110(4):e52. https://doi.org/10.1016/j.fertnstert.2018.07.161.
Yumoto K, Iwata K, Sugishima M, Yamauchi J, Nakaoka M, Tsuneto M, et al. Unstable osmolality of microdrops cultured in non-humidified incubators. J Assist Reprod Genet. 2019;36(8):1571–7. https://doi.org/10.1007/s10815-019-01515-9.
Mullen SF. Toward a predictive theoretical model for osmolality rise with non-humidified incubation: a randomized, multivariate response-surface study. Hum Reprod. 2021;36(5):1230–41. https://doi.org/10.1093/humrep/deab015.
Mestres E, Matia-Algué Q, Villamar A, Casals A, Acacio M, García-Jiménez M, et al. Characterization and comparison of commercial oils used for human embryo culture. Hum Reprod. 2021;37(2):212–25. https://doi.org/10.1093/humrep/deab245.
Shimada M, Kawano N, Terada T. Delay of nuclear maturation and reduction in developmental competence of pig oocytes after mineral oil overlay of in vitro maturation media. Reproduction. 2002;124(4):557–64. https://doi.org/10.1530/rep.0.1240557
Segers I, Adriaenssens T, Coucke W, Cortvrindt R, Smitz J. Timing of nuclear maturation and postovulatory aging in oocytes of in vitro-grown mouse follicles with or without oil overlay. Biol Reprod. 2008;78(5):859–68. https://doi.org/10.1095/biolreprod.107.062539.
Martinez CA, Nohalez A, Cuello C, Vazquez JM, Roca J, Martinez EA, et al. The use of mineral oil during in vitro maturation, fertilization, and embryo culture does not impair the developmental competence of pig oocytes. Theriogenology. 2015;83(4):693–702. https://doi.org/10.1016/j.theriogenology.2014.11.001.
Otsuki J, Nagai Y, Chiba K. Damage of embryo development caused by peroxidized mineral oil and its association with albumin in culture. Fertil Steril. 2009;91(5):1745–9. https://doi.org/10.1016/j.fertnstert.2008.03.001.
Martinez CA, Nohalez A, Ceron JJ, Rubio CP, Roca J, Cuello C, et al. Peroxidized mineral oil increases the oxidant status of culture media and inhibits in vitro porcine embryo development. Theriogenology. 2017;103:17–23. https://doi.org/10.1016/j.theriogenology.2017.07.028.
Erbach GT, Bhatnagar P, Baltz JM, Biggers JD. Zinc is a possible toxic contaminant of silicone oil in microdrop cultures of preimplantation mouse embryos. Hum Reprod. 1995;10(12):3248–54. https://doi.org/10.1093/oxfordjournals.humrep.a135897.
Martinez CA, Nohalez A, Parrilla I, Motas M, Roca J, Romero I, García-González DL, Cuello C, Rodriguez-Martinez H, Martinez EA, Gil MA. The overlaying oil type influences in vitro embryo production: differences in composition and compound transfer into incubation medium between oils. Sci Rep. 2017;7(1):10505. https://doi.org/10.1038/s41598-017-10989-5
Mestres E, Garcia-Jiménez M, Faes L, Vanrell I, Bogaert V, Jonckheere I, et al. Parameters of the Mouse Embryo Assay that affect detection of peroxides in mineral oil. Reprod Biomed Online. 2019;39(4):547–55. https://doi.org/10.1016/j.rbmo.2019.05.008.
This work was funded by the authors’ institutions.
Conflict of interest
The authors declare no competing interests.
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
About this article
Cite this article
Scarica, C., Monaco, A., Borini, A. et al. Use of mineral oil in IVF culture systems: physico-chemical aspects, management, and safety. J Assist Reprod Genet 39, 883–892 (2022). https://doi.org/10.1007/s10815-022-02479-z