Journal of Assisted Reproduction and Genetics

, Volume 35, Issue 1, pp 41–48 | Cite as

A novel fibrin-based artificial ovary prototype resembling human ovarian tissue in terms of architecture and rigidity

  • Maria Costanza Chiti
  • Marie-Madeleine Dolmans
  • Lucie Mortiaux
  • Flanco Zhuge
  • Emna Ouni
  • Parinaz Asiabi Kohneh Shahri
  • Evelyne Van Ruymbeke
  • Sophie-Demoustier Champagne
  • Jacques Donnez
  • Christiani Andrade Amorim
Fertility Preservation

Abstract

Purpose

The aim of this study is to optimize fibrin matrix composition in order to mimic human ovarian tissue architecture for human ovarian follicle encapsulation and grafting.

Methods

Ultrastructure of fresh human ovarian cortex in age-related women (n = 3) and different fibrin formulations (F12.5/T1, F30/T50, F50/T50, F75/T75), rheology of fibrin matrices and histology of isolated and encapsulated human ovarian follicles in these matrices.

Results

Fresh human ovarian cortex showed a highly fibrous and structurally inhomogeneous architecture in three age-related patients, but the mean ± SD of fiber thickness (61.3 to 72.4 nm) was comparable between patients. When the fiber thickness of four different fibrin formulations was compared with human ovarian cortex, F50/T50 and F75/T75 showed similar fiber diameters to native tissue, while F12.5/T1 was significantly different (p value < 0.01). In addition, increased concentrations of fibrin exhibited enhanced storage modulus with F50/T50, resembling physiological ovarian rigidity. Excluding F12.5/T1 from further analysis, only three remaining fibrin matrices (F30/T50, F50/T50, F75/T75) were histologically investigated. For this, frozen-thawed fragments of human ovarian tissue collected from 22 patients were used to isolate ovarian follicles and encapsulate them in the three fibrin formulations. All three yielded similar follicle recovery and loss rates soon after encapsulation. Therefore, based on fiber thickness, porosity, and rigidity, we selected F50/T50 as the fibrin formulation that best mimics native tissue.

Conclusions

Of all the different fibrin matrix concentrations tested, F50/T50 emerged as the combination of choice in terms of ultrastructure and rigidity, most closely resembling human ovarian cortex.

Keywords

Human ovarian tissue microstructure Scanning electron microscopy Fibrin matrix Porosity Isolated follicles Artificial ovary 

Notes

Acknowledgments

The authors thank Patricia Meijers for her collaboration and scientific advice on fibrin in the project. They also thank Mira Hryniuk, BA, for reviewing the English language of the manuscript and Delphine Magnin, Dolores Gonzalez, and Olivier Van Kerk for their technical assistance. This study was supported by grants from the Fonds National de la Recherche Scientifique de Belgique (FNRS) awarded to C. A. Amorim as a research associate for the FRS-FNRS and M.M. Dolmans (grant 5/4/150/5), Fonds Spéciaux de Recherche, Fondation St Luc, Foundation Against Cancer, and Wallonie-Bruxelles International, and donations from the Ferrero family.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

References

  1. 1.
    Donnez J, Dolmans MM. Ovarian cortex transplantation: 60 reported live births brings the success and worldwide expansion of the technique towards routine clinical practice. J Assist Reprod Genet. 2015;32(8):1167–70.CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    Anderson RA, Wallace WHB, Telfer EE. Ovarian tissue cryopreservation for fertility preservation: clinical and research perspectives. Human Reproduction Open. 2017;2017(1):hox001-hox.CrossRefGoogle Scholar
  3. 3.
    Jensen AK, Macklon KT, Fedder J, Ernst E, Humaidan P, Andersen CY. 86 successful births and 9 ongoing pregnancies worldwide in women transplanted with frozen-thawed ovarian tissue: focus on birth and perinatal outcome in 40 of these children. J Assist Reprod Genet. 2017;34(3):325–36.CrossRefPubMedGoogle Scholar
  4. 4.
    Smitz J, Dolmans MM, Donnez J, Fortune JE, Hovatta O, Jewgenow K, et al. Current achievements and future research directions in ovarian tissue culture, in vitro follicle development and transplantation: implications for fertility preservation. Hum Reprod Update. 2010;16(4):395–414.CrossRefPubMedPubMedCentralGoogle Scholar
  5. 5.
    Dolmans MM, Luyckx V, Donnez J, Andersen CY, Greve T. Risk of transferring malignant cells with transplanted frozen-thawed ovarian tissue. Fertil Steril. 2013;99(6):1514–22.CrossRefPubMedGoogle Scholar
  6. 6.
    Donnez J, Dolmans MM. Fertility preservation in women. Nat Rev Endocrinol. 2013;9(12):735–49.CrossRefPubMedGoogle Scholar
  7. 7.
    Soares M, Saussoy P, Maskens M, Reul H, Amorim CA, Donnez J, et al. Eliminating malignant cells from cryopreserved ovarian tissue is possible in leukaemia patients. Br J Haematol. 2017;178(2):231–239.Google Scholar
  8. 8.
    Amorim CA. Artificial ovary. In: Suzuki N, Donnez J. Gonadal tissue cryopreservation in fertility preservation. Tokyo: Springer; 2016. p. 175–192.Google Scholar
  9. 9.
    Kristensen SG, Rasmussen A, Byskov AG, Andersen CY. Isolation of pre-antral follicles from human ovarian medulla tissue. Human reproduction (Oxford, England). 2011;26(1):157–66.CrossRefGoogle Scholar
  10. 10.
    Vanacker J, Camboni A, Dath C, Van Langendonckt A, Dolmans MM, Donnez J, et al. Enzymatic isolation of human primordial and primary ovarian follicles with Liberase DH: protocol for application in a clinical setting. Fertil Steril. 2011;96(2):379–83.e3.CrossRefPubMedGoogle Scholar
  11. 11.
    Lierman S, Tilleman K, Cornelissen M, De Vos WH, Weyers S, T'Sjoen G, et al. Follicles of various maturation stages react differently to enzymatic isolation: a comparison of different isolation protocols. Reprod BioMed Online. 2015;30(2):181–90.CrossRefPubMedGoogle Scholar
  12. 12.
    Soares M, Sahrari K, Amorim CA, Saussoy P, Donnez J, Dolmans MM. Evaluation of a human ovarian follicle isolation technique to obtain disease-free follicle suspensions before safely grafting to cancer patients. Fertil Steril. 2015;104(3):672–80.e2.CrossRefPubMedGoogle Scholar
  13. 13.
    Amorim CA, Shikanov A. The artificial ovary: current status and future perspectives. Future oncology (London, England). 2016;12(20):2323–32.CrossRefGoogle Scholar
  14. 14.
    Dolmans MM, Martinez-Madrid B, Gadisseux E, Guiot Y, Yuan WY, Torre A, et al. Short-term transplantation of isolated human ovarian follicles and cortical tissue into nude mice. Reproduction (Cambridge, England). 2007;134(2):253–62.CrossRefGoogle Scholar
  15. 15.
    Vanacker J, Dolmans MM, Luyckx V, Donnez J, Amorim CA. First transplantation of isolated murine follicles in alginate. Regen Med. 2014;9(5):609–19.CrossRefPubMedGoogle Scholar
  16. 16.
    Luyckx V, Dolmans MM, Vanacker J, Legat C, Fortuno Moya C, Donnez J, et al. A new step toward the artificial ovary: survival and proliferation of isolated murine follicles after autologous transplantation in a fibrin scaffold. Fertil Steril. 2014;101(4):1149–56.CrossRefPubMedGoogle Scholar
  17. 17.
    Kniazeva E, Hardy AN, Boukaidi SA, Woodruff TK, Jeruss JS, Shea LD. Primordial follicle transplantation within designer biomaterial grafts produce live births in a mouse infertility model. Sci Rep. 2015;5:17709.CrossRefPubMedPubMedCentralGoogle Scholar
  18. 18.
    Laronda MM, Jakus AE, Whelan KA, Wertheim JA, Shah RN, Woodruff TK. Initiation of puberty in mice following decellularized ovary transplant. Biomaterials. 2015;50:20–9.CrossRefPubMedPubMedCentralGoogle Scholar
  19. 19.
    Laronda MM, Rutz AL, Xiao S, Whelan KA, Duncan FE, Roth EW, et al. A bioprosthetic ovary created using 3D printed microporous scaffolds restores ovarian function in sterilized mice. Nat Commun. 2017;8:15261.CrossRefPubMedPubMedCentralGoogle Scholar
  20. 20.
    Kim J, Perez AS, Claflin J, David A, Zhou H, Shikanov A. Synthetic hydrogel supports the function and regeneration of artificial ovarian tissue in mice. Npj Regenerative Medicine. 2016;1:16010.CrossRefPubMedPubMedCentralGoogle Scholar
  21. 21.
    Paulini F, Vilela JM, Chiti MC, Donnez J, Jadoul P, Dolmans MM, et al. Survival and growth of human preantral follicles after cryopreservation of ovarian tissue, follicle isolation and short-term xenografting. Reprod BioMed Online. 2016;33(3):425–32.CrossRefPubMedGoogle Scholar
  22. 22.
    Chiti MC, Dolmans MM, Donnez J, Amorim CA. Fibrin in reproductive tissue engineering: a review on its application as a biomaterial for fertility preservation. Ann Biomed Eng. 2017;45(7):1650–1663.Google Scholar
  23. 23.
    Chiti MC, Dolmans MM, Orellana R, Soares M, Paulini F, Donnez J, et al. Influence of follicle stage on artificial ovary outcome using fibrin as a matrix. Human reproduction (Oxford, England). 2016;31(2):427–35.Google Scholar
  24. 24.
    Chiti MC, Dolmans MM, Lucci CM, Paulini F, Donnez J, Amorim CA. Further insights into the impact of mouse follicle stage on graft outcome in an artificial ovary environment. Mol Hum Reprod. 2017;23(6):381–392.Google Scholar
  25. 25.
    Amorim CA. The ovarian follicle microenviroment and the artificial ovary. Shanghai: 4th World Congress of International Society for Fertility Preservation; 2015.Google Scholar
  26. 26.
    Chiti MC. A modified and tailored human follicle isolation procedure improve follicle survival in an artificial ovary prototype after short term xenografting. J Ovarian Res. 2017;10(1):71.Google Scholar
  27. 27.
    Fox CH, Johnson FB, Whiting J, Roller PP. Formaldehyde fixation. The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society. 1985;33(8):845–53.CrossRefGoogle Scholar
  28. 28.
    Smith RM, Shikanov A, Kniazeva E, Ramadurai D, Woodruff TK, Shea LD. Fibrin-mediated delivery of an ovarian follicle pool in a mouse model of infertility. Tissue Eng A. 2014;20(21–22):3021–30.CrossRefGoogle Scholar
  29. 29.
    Rodgers RJ, Irving-Rodgers HF, Russell DL. Extracellular matrix of the developing ovarian follicle. Reproduction (Cambridge, England). 2003;126(4):415–24.CrossRefGoogle Scholar
  30. 30.
    Cho A, Howell VM, Colvin EK. The extracellular matrix in epithelial ovarian cancer—a piece of a puzzle. Front Oncol. 2015;5:245.CrossRefPubMedPubMedCentralGoogle Scholar
  31. 31.
    Herraiz S, Diaz-Garcia C, Pellicer A. Ovarian tissue cryopreservation: slow freezing. In: Suzuki N, Donnez J, editors. Gonadal tissue cryopreservation in fertility preservation. Tokyo: Springer Japan; 2016. p. 53–77.Google Scholar
  32. 32.
    Armstrong CG, Mow VC. Variations in the intrinsic mechanical properties of human articular cartilage with age, degeneration, and water content. J Bone Joint Surg Am. 1982;64(1):88–94.CrossRefPubMedGoogle Scholar
  33. 33.
    Chaffin CL, Vandevoort CA. Follicle growth, ovulation, and luteal formation in primates and rodents: a comparative perspective. Experimental biology and medicine (Maywood, NJ). 2013;238(5):539–48.CrossRefGoogle Scholar
  34. 34.
    Wolberg AS. Thrombin generation and fibrin clot structure. Blood Rev. 2007;21(3):131–42.CrossRefPubMedGoogle Scholar
  35. 35.
    Kim BS, Sung HM, You HK, Lee J. Effects of fibrinogen concentration on fibrin glue and bone powder scaffolds in bone regeneration. J Biosci Bioeng. 2014;118(4):469–75.CrossRefPubMedGoogle Scholar
  36. 36.
    Germain L, De Berdt P, Vanacker J, Leprince J, Diogenes A, Jacobs D, et al. Fibrin hydrogels to deliver dental stem cells of the apical papilla for regenerative medicine. Regen Med. 2015;10(2):153–67.CrossRefPubMedGoogle Scholar
  37. 37.
    Woodruff TK, Shea LD. A new hypothesis regarding ovarian follicle development: ovarian rigidity as a regulator of selection and health. J Assist Reprod Genet. 2011;28(1):3–6.CrossRefPubMedGoogle Scholar
  38. 38.
    Choi JK, Agarwal P, Huang H, Zhao S, He X. The crucial role of mechanical heterogeneity in regulating follicle development and ovulation with engineered ovarian microtissue. Biomaterials. 2014;35(19):5122–8.CrossRefPubMedPubMedCentralGoogle Scholar
  39. 39.
    Wood CD, Vijayvergia M, Miller FH, Carroll T, Fasanati C, Shea LD, et al. Multi-modal magnetic resonance elastography for noninvasive assessment of ovarian tissue rigidity in vivo. Acta Biomater. 2015;13:295–300.CrossRefPubMedGoogle Scholar
  40. 40.
    Duong H, Wu B, Tawil B. Modulation of 3D fibrin matrix stiffness by intrinsic fibrinogen-thrombin compositions and by extrinsic cellular activity. Tissue Eng A. 2009;15(7):1865–76.CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2017

Authors and Affiliations

  • Maria Costanza Chiti
    • 1
  • Marie-Madeleine Dolmans
    • 1
    • 2
  • Lucie Mortiaux
    • 3
  • Flanco Zhuge
    • 3
  • Emna Ouni
    • 1
  • Parinaz Asiabi Kohneh Shahri
    • 1
  • Evelyne Van Ruymbeke
    • 3
  • Sophie-Demoustier Champagne
    • 3
  • Jacques Donnez
    • 4
  • Christiani Andrade Amorim
    • 1
  1. 1.Pôle de Recherche en Gynécologie, Institut de Recherche Expérimentale et CliniqueUniversité Catholique de LouvainBrusselsBelgium
  2. 2.Gynecology DepartmentCliniques Universitaires Saint-LucBrusselsBelgium
  3. 3.Institute of Condensed Matter and Nanosciences, Bio and Soft MatterUniversité Catholique de LouvainLouvain-la-NeuveBelgium
  4. 4.Society for Research into InfertilityBrusselsBelgium

Personalised recommendations