Lower follicular n-3 polyunsaturated fatty acid levels are associated with a better response to ovarian stimulation

  • José-Ignacio Ruiz-Sanz
  • Irantzu Pérez-Ruiz
  • Susana Meijide
  • Marcos Ferrando
  • Zaloa Larreategui
  • María-Begoña Ruiz-LarreaEmail author
Assisted Reproduction Technologies



To analyze in detail the fatty acid (FA) composition of follicular fluid (FF) from two-sized follicles at oocyte retrieval and to determine associations of the FAs from large follicles with the woman’s age and the response to ovarian stimulation.


Observational study.


University and fertility clinic.


Sixty-four women (age 19–46), consisting of unfertile patients and oocyte donors, undergoing controlled ovarian stimulation.



Main outcome measure(s)

FF from small (< 12 mm) and large (≥ 18 mm) follicles was collected at oocyte retrieval. FAs by gas chromatography-flame ionization detection.


Thirty-two FAs with chain lengths ranging from 14 to 25 carbons were identified. There was a readjustment in FA distribution as follicle size increased, raising very long-chain saturated FAs, nervonic (24:1n-9), arachidonic (20:4n-6), and n-3 polyunsaturated FAs (PUFA, P < 0.001), the latter mainly due to an increase in docosahexaenoic acid (22:6n-3, DHA). In large follicles, double bond and peroxidizability indices and total n-3 PUFA, particularly DHA, correlated positively with the woman’s age and negatively with the number of total and mature oocytes, total and top-quality embryos, and fertilization rate.


We have described 32 FAs in ovarian FF, of which 16 changed their distribution with follicle size. The results also indicate that lower n-3 PUFA levels in large follicles, which are associated with younger women, predict a better response to ovarian stimulation based on the recovery of total and mature oocytes, total and top-quality embryos, and fertilization rate per cycle.

Key message

The fatty acid profile of ovarian FF changes as the follicle grows and lower n-3 PUFA levels in large follicles, associated with younger women, predict a better response to ovarian stimulation.


Female infertility Fatty acids Aging Gas chromatography Follicular fluid 



Supported by the “PN de I+D+I” of the Spanish Ministry of Science and Innovation, “ISCIII-Subdirección General de Evaluación y Fomento de la Investigación” and FEDER (ref. FIS/FEDER PI11/02559), University of the Basque Country UPV/EHU (ref. GIU16/62), and the Basque Government (Department of Education, Universities and Research, predoctoral grant to IP, and Department of Development, Economy and Competitiveness, SPRI, ref. IG-2013 0001214).

Compliance with ethical standards

Ethical approval

The Ethics Committee of the University UPV/EHU (Ethics Committee for Research involving Human Subjects, CEISH) approved the human subject protocol (CEISH/96/2011/RUIZLARREA), and the study was performed according to the UPV/EHU and IVI-RMA Bilbao agreement, Ref. 2012/01. The project complies with the Spanish Law of Assisted Reproductive Technologies (14/2006). Written informed consent was obtained from all trial subjects for participation in the study.

Conflict of interest

The authors declare that they have no conflict of interest.


  1. 1.
    ESHRE Capri Workshop Group. A prognosis-based approach to infertility: understanding the role of time. Hum Reprod. 2017;32:1556–9.CrossRefGoogle Scholar
  2. 2.
    Sutton ML, Gilchrist RB, Thompson JG. Effects of in-vivo and in-vitro environments on the metabolism of the cumulus-oocyte complex and its influence on oocyte developmental capacity. Hum Reprod Update. 2003;9:35–48.CrossRefGoogle Scholar
  3. 3.
    Schmitz G, Ecker J. The opposing effects of n-3 and n-6 fatty acids. Prog Lipid Res. 2008;47:147–55 Review.CrossRefGoogle Scholar
  4. 4.
    Wathes DC, Abayasekara DR, Aitken RJ. Polyunsaturated fatty acids in male and female reproduction. Biol Reprod. 2007;77:190–201.CrossRefGoogle Scholar
  5. 5.
    Jungheim ES, Macones GA, Odem RR, Patterson BW, Lanzendorf SE, Ratts VS, et al. Associations between free fatty acids, cumulus oocyte complex morphology and ovarian function during in vitro fertilization. Fertil Steril. 2011;95:1970–4.CrossRefGoogle Scholar
  6. 6.
    Valckx SD, Arias-Alvarez M, De Pauw I, Fievez V, Vlaeminck B, Fransen E, et al. Fatty acid composition of the follicular fluid of normal weight, overweight and obese women undergoing assisted reproductive treatment: a descriptive cross-sectional study. Reprod Biol Endocrinol. 2014;12:13. Scholar
  7. 7.
    Soares SR, Troncoso C, Bosch E, Serra V, Simón C, Remohí J, et al. Age and uterine receptiveness: predicting the outcome of oocyte donation cycles. J Clin Endocrinol Metab. 2005;90:4399–404.CrossRefGoogle Scholar
  8. 8.
    Faul F, Erdfelder E, Lang AG, Buchner A. G*power 3: a flexible statistical power analysis program for the social, behavioral, and biomedical sciences. Behav Res Methods. 2007;39:175–91.CrossRefGoogle Scholar
  9. 9.
    ASEBIR. Criterios ASEBIR de valoración morfológica de oocitos, embriones tempranos y blastocistos humanos. Cuadernos de Embriología Clínica. 3rd ed. Glóbalo, Agencia Creativa Digital, Madrid; 2015. ISSN: 1888–8011.Google Scholar
  10. 10.
    Lepage, Roy CC. Direct transesterification of all classes of lipids in one step reaction. J Lipid Res. 1986;27:114–20.PubMedGoogle Scholar
  11. 11.
    Shaaker M, Rahimipour A, Nouri M, Khanaki K, Darabi M, Farzadi L, et al. Fatty acid composition of human follicular fluid phospholipids and fertilization rate in assisted reproductive techniques. Iran Biomed J. 2012;16:162–8.PubMedPubMedCentralGoogle Scholar
  12. 12.
    Pantasri T, Wu LL, Hull ML, Sullivan TR, Barry M, Norman RJ, et al. Distinct localisation of lipids in the ovarian follicular environment. Reprod Fertil Dev. 2015;27:593. Scholar
  13. 13.
    Warzych E, Pawlak P, Pszczola M, Cieslak A, Madeja ZE, Lechniak D. Interactions of bovine oocytes with follicular elements with respect to lipid metabolism. Anim Sci J. 2017;55:1491–7.CrossRefGoogle Scholar
  14. 14.
    Homa ST, Brown CA. Changes in linoleic acid during follicular development and inhibition of spontaneous breakdown of germinal vesicles in cumulus-free bovine oocytes. J Reprod Fertil. 1992;94:153–60.CrossRefGoogle Scholar
  15. 15.
    Marei WF, Wathes DC, Fouladi-Nashta AA. Impact of linoleic acid on bovine oocyte maturation and embryo development. Reproduction. 2010;139:979–88.CrossRefGoogle Scholar
  16. 16.
    Ghaffarilaleh V, Fouladi-Nashta A, Paramio MT. Effect of α-linolenic acid on oocyte maturation and embryo development of prepubertal sheep oocytes. Theriogenology. 2014;82:686–96.CrossRefGoogle Scholar
  17. 17.
    Nonogaki T, Noda Y, Goto Y, Kishi J, Mori T. Developmental blockage of mouse embryos caused by fatty acids. J Assist Reprod Genet. 1994;11:482–8.CrossRefGoogle Scholar
  18. 18.
    Ciepiela P, Bączkowski T, Drozd A, Kazienko A, Stachowska E, Kurzawa R. Arachidonic and linoleic acid derivatives impact oocyte ICSI fertilization--a prospective analysis of follicular fluid and a matched oocyte in a 'one follicle--one retrieved oocyte--one resulting embryo' investigational setting. PLoS One. 2015;10:e0119087. Scholar
  19. 19.
    Wiener-Megnazi Z, Vardi L, Lissak A, Shnizer S, Reznick AZ, Ishai D, et al. Oxidative stress indices in follicular fluid as measured by the thermochemiluminescence assay correlate with outcome parameters in in vitro fertilization. Fertil Steril. 2004;82:1171–6.CrossRefGoogle Scholar
  20. 20.
    Shkolnik K, Tadmor A, Ben-Dor S, Nevo N, Galiani D, Dekel N. Reactive oxygen species are indispensable in ovulation. Proc Natl Acad Sci U S A. 2011;108:1462–7.CrossRefGoogle Scholar
  21. 21.
    Espey LL, Stein VI, Dumitrescu J. Survey of antiinflammatory agents and related drugs as inhibitors of ovulation in the rabbit. Fertil Steril. 1982;38:238–47.CrossRefGoogle Scholar
  22. 22.
    Bolton-Smith C, Woodward M, Tavendale R. Evidence for age-related differences in the fatty acid composition of human adipose tissue, independent of diet. Eur J Clin Nutr. 1997;51:619–24.CrossRefGoogle Scholar
  23. 23.
    Crowe FL, Skeaff CM, Green TJ, Gray AR. Serum n-3 long-chain PUFA differ by sex and age in a population-based survey of New Zealand adolescents and adults. Br J Nutr. 2008;99:168–74.CrossRefGoogle Scholar
  24. 24.
    Walker CG, Browning LM, Mander AP, Madden J, West AL, Calder PC, et al. Age and sex differences in the incorporation of EPA and DHA into plasma fractions, cells and adipose tissue in humans. Br J Nutr. 2014;111:679–89.CrossRefGoogle Scholar
  25. 25.
    Borsonelo EC, Galduróz JCF. The role of polyunsaturated fatty acids (PUFAs) in development, aging and substance abuse disorders: review and propositions. Prostaglandins Leukot Essent Fatty Acids. 2008;78:237–45.CrossRefGoogle Scholar
  26. 26.
    Li Y, Kang JX, Leaf A. Differential effects of various eicosanoids on the production or prevention of arrhythmias in cultured neonatal rat cardiac myocytes. Prostaglandins. 1997;54:511–30.CrossRefGoogle Scholar
  27. 27.
    McKeegan PJ, Sturmey RG. The role of fatty acids in oocyte and early embryo development. Reprod Fertil Dev. 2011;24:59–67.CrossRefGoogle Scholar
  28. 28.
    Jové M, Naudí A, Gambini J, Borras C, Cabré R, Portero-Otín M, et al. A stress-resistant Lipidomic signature confers extreme longevity to humans. J Gerontol A Biol Sci Med Sci. 2017;72:30–7.Google Scholar
  29. 29.
    Otsuka R, Kato Y, Imai T, Ando F, Shimokata H. Higher serum EPA or DHA, and lower ARA compositions with age independent fatty acid intake in Japanese aged 40 to 79. Lipids. 2013;48:719–27.CrossRefGoogle Scholar
  30. 30.
    Otsuka R, Kato Y, Imai T, Ando F, Shimokata H. Secular trend of serum docosahexaenoic acid, eicosapentaenoic acid, and arachidonic acid concentrations among Japanese. A 4- and13-year descriptive epidemiologic study. Prostaglandins Leukot Essent Fatty Acids. 2015;94:35–42.CrossRefGoogle Scholar
  31. 31.
    Rajalahti T, Lin C, Mjøs SA, Kvalheim OM. Changes in serum fatty acid and lipoprotein subclass concentrations from prepuberty to adulthood and during aging. Metabolomics. 2016;12:51.CrossRefGoogle Scholar
  32. 32.
    Risé P, Tragni E, Ghezzi S, Agostoni C, Marangoni F, Poli A, et al. IDEFICS consortium., CHECK group. Different patterns characterize omega 6 and omega 3 long chain polyunsaturated fatty acid levels in blood from Italian infants, children, adults and elderly. Prostaglandins Leukot Essent Fatty Acids. 2013;89:215–20.CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Free Radicals and Oxidative Stress (FROS) Research Group of the Department of Physiology, Medicine and Nursing SchoolUniversity of the Basque Country UPV/EHULeioaSpain
  2. 2.BioCruces Health Research InstituteBarakaldoSpain
  3. 3.Valencian Institute of Infertility (IVI-RMA)-BilbaoLeioaSpain

Personalised recommendations