Journal of Assisted Reproduction and Genetics

, Volume 35, Issue 8, pp 1407–1417 | Cite as

Presence of growth/differentiation factor-15 cytokine in human follicular fluid, granulosa cells, and oocytes

  • Karel SoučekEmail author
  • Alice Malenovská
  • Zuzana Kahounová
  • Ján Remšík
  • Zuzana Holubcová
  • Tomáš Soukup
  • Daniela Kurfürstová
  • Jan Bouchal
  • Tereza Suchánková
  • Eva Slabáková
  • Aleš HamplEmail author
Reproductive Physiology and Disease



The purpose of the study was to determine whether the GDF-15 is present in follicular fluid; to evaluate if there is a relation between follicular and serum levels of GDF-15 and fertility status of study subjects; and to test whether granulosa cells, oocytes, or both produce GDF-15.


This study used follicular fluid (FF, serum, and oocytes obtained under informed consent from women undergoing oocyte retrieval for in vitro fertilization. It also used ovaries from deceased preterm newborns. Collection of FF and blood at the time of oocyte retrieval, ELISA and western blot were performed to determine levels and forms of GDF-15. Concentrations of GDF-15 in FF and serum, its expression in ovarian tissue, and secretion from granulosa cells were analyzed.


GDF-15 concentration in FF ranged from 35 to 572 ng/ml, as determined by ELISA. Western blot analysis revealed the GDF-15 pro-dimer only in FF. Both normal healthy and cancerous granulosa cells secreted GDF-15 into culture media. Primary oocytes displayed cytoplasmic GDF-15 positivity in immunostained newborn ovaries, and its expression was also observed in fully grown human oocytes.


To the best of our knowledge, this is the first documentation of cytokine GDF-15 presence in follicular fluid. Its concentration was not associated with donor/patient fertility status. Our data also show that GDF-15 is expressed and inducible in both normal healthy and cancerous granulosa cells, as well as in oocytes.


Follicular fluid Growth/differentiation factor-15 Follicular granulosa cells IVF 



The authors would like to thank Iva Lišková, Martina Urbánková, and Kateřina Svobodová for their superb technical assistance; Petra Ovesná for helping with statistical analyses, Monika Smějová, and Veronika Toporcerová for helping with protein quantifications and for western blot analyses respectively.

Author’s roles

KS carried out ELISA procedures and conceptualized the study, contributing to its design, coordination, and manuscript draft. Tsu and JR performed western blot testing; ZK and ES conducted qRT-PCR analysis, and IHC analysis was undertaken by JB and DK. TSo established the primary granulosa cell lines. AM and ZH completed sample preparation. AH participated in study design and with ES helped to draft the manuscript. All authors have read and approved the final manuscript.


This work was supported by the following grants: 15-11707S from Czech Science Foundation, by Ministry of Health of the Czech Republic, grant no. 15-33999A, all rights reserved (KS); project PROGRES Q40/06 (Charles University [TSo]); project mobility no. 7AMB16AT022 from the Ministry of Education, Youth and Sports; project HistoPARK (CZ.1.07/2.3.00/20.0185, [KS, AH]), CELLBIOL (CZ.1.07/2.3.00/30.0030 [TS]); project no. LQ1605 from the National Program of Sustainability II (MEYS CR; KS, AH); project no. LO1304 from the National Program of Sustainability I (MEYS CR; JB, DK); and the European Union—project ICRC-ERA-HumanBridge (no. 316345; KS, AH).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

10815_2018_1230_MOESM1_ESM.pdf (653 kb)
ESM 1 Supplementary Fig. 1 Flow-chart diagram. Supplementary Fig. 2 Validation of the anti-GDF15 antibody was performed on formalin-fixed paraffin-embedded cell lines with known expression of GDF15. High (LNCaP cell line) and low expression (BPH1 cell line) of GDF15 was detected both by immunohistochemistry and western blotting. Magnification 200X, scale bars represent 50 μm. Supplementary Fig. 3 Immunohistochemical staining of HIF-1α in the ovary from the preterm newborn 2. The staining pattern of some follicles is different from the preterm newborn 1, probably due to longer time to autopsy (the autopsies were performed 28 and 47 h after exitus of newborns 1 and 2, respectively). Importantly, the staining pattern of both HIF-1α and GDF15 in the cytoplasm of oocytes is similar in both cases (please compare to the Fig. 3). Magnification 400X, scale bar represents 20 μm. Supplementary Table 1. Results of T-test of donor vs. patient characteristics. (PDF 652 kb)


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Copyright information

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

Authors and Affiliations

  1. 1.Institute of Biophysics of the Czech Academy of SciencesBrnoCzech Republic
  2. 2.International Clinical Research CenterSt. Anne’s University Hospital BrnoBrnoCzech Republic
  3. 3.Reprofit International Clinic of Reproductive MedicineBrnoCzech Republic
  4. 4.Department of Experimental Biology, Faculty of ScienceMasaryk UniversityBrnoCzech Republic
  5. 5.Department of Histology and Embryology, Faculty of MedicineMasaryk UniversityBrnoCzech Republic
  6. 6.Faculty of Medicine in Hradec Králové, Department of Histology and EmbryologyCharles University in PragueHradec KrálovéCzech Republic
  7. 7.Department of Clinical and Molecular Pathology, Institute of Molecular and Translational Medicine, Faculty of Medicine and DentistryPalacky University OlomoucOlomoucCzech Republic

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