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.
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.
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.
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.
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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.
Anderson RA, Wallace WHB, Telfer EE. Ovarian tissue cryopreservation for fertility preservation: clinical and research perspectives. Human Reproduction Open. 2017;2017(1):hox001-hox.
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.
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.
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.
Donnez J, Dolmans MM. Fertility preservation in women. Nat Rev Endocrinol. 2013;9(12):735–49.
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.
Amorim CA. Artificial ovary. In: Suzuki N, Donnez J. Gonadal tissue cryopreservation in fertility preservation. Tokyo: Springer; 2016. p. 175–192.
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.
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.
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.
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.
Amorim CA, Shikanov A. The artificial ovary: current status and future perspectives. Future oncology (London, England). 2016;12(20):2323–32.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
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.
Amorim CA. The ovarian follicle microenviroment and the artificial ovary. Shanghai: 4th World Congress of International Society for Fertility Preservation; 2015.
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.
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.
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.
Rodgers RJ, Irving-Rodgers HF, Russell DL. Extracellular matrix of the developing ovarian follicle. Reproduction (Cambridge, England). 2003;126(4):415–24.
Cho A, Howell VM, Colvin EK. The extracellular matrix in epithelial ovarian cancer—a piece of a puzzle. Front Oncol. 2015;5:245.
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.
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.
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.
Wolberg AS. Thrombin generation and fibrin clot structure. Blood Rev. 2007;21(3):131–42.
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.
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.
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.
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.
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.
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.
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.
Conflict of interest
The authors declare that they have no conflict of interest.
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Chiti, M.C., Dolmans, MM., Mortiaux, L. et al. A novel fibrin-based artificial ovary prototype resembling human ovarian tissue in terms of architecture and rigidity. J Assist Reprod Genet 35, 41–48 (2018). https://doi.org/10.1007/s10815-017-1091-3
- Human ovarian tissue microstructure
- Scanning electron microscopy
- Fibrin matrix
- Isolated follicles
- Artificial ovary