CoChat
Sign Up
Cited by 72Tissue Engineering and Regenerative Medicine3D Printing in Biomedical ResearchSperm and Testicular Function

Development of a Cytocompatible Scaffold from Pig Immature Testicular Tissue Allowing Human Sertoli Cell Attachment, Proliferation and Functionality

Open access
Vermeulen, Maxime, Del Vento, Federico, De Michele, Francesca, Poels, Jonathan, Wyns, Christine·2018·International Journal of Molecular Sciences
DOI 10.3390/ijms19010227PMC PMC5796176
Open this paper in CoChat — free accountOpen PDF

Abstract

Cryopreservation of immature testicular tissue before chemo/radiotherapy is the only option to preserve fertility of cancer-affected prepubertal boys. To avoid reintroduction of malignant cells, development of a transplantable scaffold by decellularization of pig immature testicular tissue (ITT) able to support decontaminated testicular cells could be an option for fertility restoration in these patients. We, therefore, compared decellularization protocols to produce a cytocompatible scaffold. Fragments of ITT from 15 piglets were decellularized using three protocols: sodium dodecyl sulfate (SDS)-Triton (ST), Triton-SDS-Triton (TST) and trypsin 0.05%/ethylenediaminetetraacetic acid (EDTA) 0.02%-Triton (TET) with varying detergent concentrations. All protocols were able to lower DNA levels. Collagen retention was demonstrated in all groups except ST 1%, and a significant decrease in glycosaminoglycans was observed in the TST 1% and TET 1% groups. When Sertoli cells (SCs) were cultured with decellularized tissue, no signs of cytotoxicity were detected. A higher SC proliferation rate and greater stem cell factor secretion were observed than with SCs cultured without scaffold. ST 0.01% and TET 3% conditions offered the best compromise in terms of DNA elimination and extracellular matrix (ECM) preservation, while ensuring good attachment, proliferation and functionality of human SCs. This study demonstrates the potential of using decellularized pig ITT for human testicular tissue engineering purposes.

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Integrity signals

Registry

Crossref · 10.3390/ijms19010227

Cites

Statistics

Claims the paper makes

The claims and statistics of this paper haven’t been checked yet.

Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.

A few free checks a day · papers already verified open instantly · results are public

Claims other papers make about this one

Each is checked against this paper’s own text: does it say what the citing paper says it does?

  • Vermeulen et al. developed hydrogels from decellularized porcine immature testicular tissue (ITT) that successfully generated testicular organoids (TOs).

    “Vermeulen et al. developed hydrogels from decellularized porcine immature testicular tissue (ITT) that successfully generated testicular organoids (TOs).”

    From Bioactive scaffolds for tissue engineering: A review of decellularized extracellular matrix applications and innovations (2025) · cites this paper as [200]

    The Abstract reports a decellularized pig immature testicular tissue scaffold that supported human Sertoli cells, not a hydrogel that generated organoids. The Discussion explicitly proposes developing a hydrogel as future work and describes organoid formation as an application reported elsewhere. (LLM, conf=0.91)

    Checked by a reasoning model — may be wrong.

    Experimental — accuracy not yet measured

  • They demonstrated that growth factors in TOs derived from decellularized ITT were better preserved, indicating potential for enhanced restoration of reproductive capability.

    “They demonstrated that growth factors in TOs derived from decellularized ITT were better preserved, indicating the potential for enhanced restoration of reproductive capability.[ 200 ]”

    From Bioactive scaffolds for tissue engineering: A review of decellularized extracellular matrix applications and innovations (2025) · cites this paper as [200]

    Results §2.2 reports preservation of extracellular-matrix components in decellularized immature testicular tissue, and the Discussion reports stem cell factor secretion by Sertoli cells cultured on the scaffolds. The paper does not demonstrate better preservation of growth factors in testicular organoids (TOs); it discusses organoid formation as a future possibility, not an outcome of this study. Fertility restoration is likewise presented as a potential application, not a demonstrated improvement. (LLM, conf=0.86)

    Experimental — accuracy not yet measured

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

The self-check needs the paper’s full text.

The citation lists haven’t been fetched yet

References 47 · Cited by 72

Research works printed but not linked: 43

  1. [1]Gatta G. Botta L. Rossi S. Aareleid T. Bielska-Lasota M. Clavel J. Dimitrova N. Jakab Z. Kaatsch P. Lacour B. Childhood cancer survival in Europe 1999–2007: Results of eurocare-5—A population-based study Lancet Oncol. 2014 15 35 47 10.1016/S1470-2045(13)70548-5 24314616 · DOI
  2. [2]Wyns C. Curaba M. Petit S. Vanabelle B. Laurent P. Wese J.F. Donnez J. Management of fertility preservation in prepubertal patients: 5 years’ experience at the catholic university of louvain Hum. Reprod. 2011 26 737 747 10.1093/humrep/deq387 21227939 · DOI
  3. [3]Wyns C. Collienne C. Shenfield F. Robert A. Laurent P. Roegiers L. Brichard B. Fertility preservation in the male pediatric population: Factors influencing the decision of parents and children Hum. Reprod. 2015 30 2022 2030 10.1093/humrep/dev161 26141713 · DOI
  4. [4]Sato T. Katagiri K. Gohbara A. Inoue K. Ogonuki N. Ogura A. Kubota Y. Ogawa T. In vitro production of functional sperm in cultured neonatal mouse testes Nature 2011 471 504 507 10.1038/nature09850 21430778 · DOI
  5. [5]Liu Z. Nie Y.H. Zhang C.C. Cai Y.J. Wang Y. Lu H.P. Li Y.Z. Cheng C. Qiu Z.L. Sun Q. Generation of macaques with sperm derived from juvenile monkey testicular xenografts Cell Res. 2016 26 139 142 10.1038/cr.2015.112 26369429 PMC4816125 · DOI
  6. [6]Brinster R.L. Avarbock M.R. Germline transmission of donor haplotype following spermatogonial transplantation Proc. Natl. Acad. Sci. USA 1994 91 11303 11307 10.1073/pnas.91.24.11303 7972054 PMC45219 · DOI
  7. [7]Hamra F.K. Gatlin J. Chapman K.M. Grellhesl D.M. Garcia J.V. Hammer R.E. Garbers D.L. Production of transgenic rats by lentiviral transduction of male germ-line stem cells Proc. Natl. Acad. Sci. USA 2002 99 14931 14936 10.1073/pnas.222561399 12391306 PMC137522 · DOI
  8. [8]Honaramooz A. Behboodi E. Megee S.O. Overton S.A. Galantino-Homer H. Echelard Y. Dobrinski I. Fertility and germline transmission of donor haplotype following germ cell transplantation in immunocompetent goats Biol. Reprod. 2003 69 1260 1264 10.1095/biolreprod.103.018788 12801978 · DOI
  9. [9]Trefil P. Micakova A. Mucksova J. Hejnar J. Poplstein M. Bakst M.R. Kalina J. Brillard J.P. Restoration of spermatogenesis and male fertility by transplantation of dispersed testicular cells in the chicken Biol. Reprod. 2006 75 575 581 10.1095/biolreprod.105.050278 16807385 · DOI
  10. [10]Herrid M. Olejnik J. Jackson M. Suchowerska N. Stockwell S. Davey R. Hutton K. Hope S. Hill J.R. Irradiation enhances the efficiency of testicular germ cell transplantation in sheep Biol. Reprod. 2009 81 898 905 10.1095/biolreprod.109.078279 19571259 · DOI
  11. [11]Kawasaki T. Saito K. Sakai C. Shinya M. Sakai N. Production of zebrafish offspring from cultured spermatogonial stem cells Genes Cells 2012 17 316 325 10.1111/j.1365-2443.2012.01589.x 22390480 · DOI
  12. [12]Hermann B.P. Sukhwani M. Winkler F. Pascarella J.N. Peters K.A. Sheng Y. Valli H. Rodriguez M. Ezzelarab M. Dargo G. Spermatogonial stem cell transplantation into rhesus testes regenerates spermatogenesis producing functional sperm Cell Stem Cell 2012 11 715 726 10.1016/j.stem.2012.07.017 23122294 PMC3580057 · DOI
  13. [13]Picton H.M. Wyns C. Anderson R.A. Goossens E. Jahnukainen K. Kliesch S. Mitchell R.T. Pennings G. Rives N. Tournaye H. A european perspective on testicular tissue cryopreservation for fertility preservation in prepubertal and adolescent boys Hum. Reprod. 2015 30 2463 2475 10.1093/humrep/dev190 26358785 · DOI
  14. [14]Ginsberg J.P. Carlson C.A. Lin K. Hobbie W.L. Wigo E. Wu X. Brinster R.L. Kolon T.F. An experimental protocol for fertility preservation in prepubertal boys recently diagnosed with cancer: A report of acceptability and safety Hum. Reprod. 2010 25 37 41 10.1093/humrep/dep371 19861330 PMC2794668 · DOI
  15. [15]Ginsberg J.P. Li Y. Carlson C.A. Gracia C.R. Hobbie W.L. Miller V.A. Mulhall J. Shnorhavorian M. Brinster R.L. Kolon T.F. Testicular tissue cryopreservation in prepubertal male children: An analysis of parental decision-making Pediatr. Blood Cancer 2014 61 1673 1678 10.1002/pbc.25078 24777742 PMC4676076 · DOI
  16. [16]Davidoff M.S. Breucker H. Holstein A.F. Seidl K. Cellular architecture of the lamina propria of human seminiferous tubules Cell Tissue Res. 1990 262 253 261 10.1007/BF00309880 2076533 · DOI
  17. [17]Kerr J. Loveland K. O’Bryan M. de Kretser D. Cytology of the testis and intrinsic control mechanisms Knobil and Neill’s Physiology of Reproduction Elsevier Amsterdam, The Netherlands 2006 Volume 1 827 947
  18. [18]Skinner M.K. Tung P.S. Fritz I.B. Cooperativity between sertoli cells and testicular peritubular cells in the production and deposition of extracellular matrix components J. Cell Biol. 1985 100 1941 1947 10.1083/jcb.100.6.1941 3889013 PMC2113598 · DOI
  19. [19]Tung P.S. Skinner M.K. Fritz I.B. Cooperativity between sertoli cells and peritubular myoid cells in the formation of the basal lamina in the seminiferous tubule Ann. N. Y. Acad. Sci. 1984 438 435 446 10.1111/j.1749-6632.1984.tb38304.x 6598327 · DOI
  20. [20]Tung P.S. Fritz I.B. Morphogenetic restructuring and formation of basement membranes by sertoli cells and testis peritubular cells in co-culture: Inhibition of the morphogenetic cascade by cyclic amp derivatives and by blocking direct cell contact Dev. Biol. 1987 120 139 153 10.1016/0012-1606(87)90112-6 2434375 · DOI
  21. [21]Hadley M.A. Byers S.W. Suarez-Quian C.A. Kleinman H.K. Dym M. Extracellular matrix regulates sertoli cell differentiation, testicular cord formation, and germ cell development in vitro J. Cell Biol. 1985 101 1511 1522 10.1083/jcb.101.4.1511 4044644 PMC2113921 · DOI
  22. [22]Cheng C.Y. Wong E.W. Yan H.H. Mruk D.D. Regulation of spermatogenesis in the microenvironment of the seminiferous epithelium: New insights and advances Mol. Cell. Endocrinol. 2010 315 49 56 10.1016/j.mce.2009.08.004 19682538 PMC3516447 · DOI
  23. [23]Pollanen P.P. Kallajoki M. Risteli L. Risteli J. Suominen J.J. Laminin and type iv collagen in the human testis Int. J. Androl. 1985 8 337 347 10.1111/j.1365-2605.1985.tb00846.x 3912331 · DOI
  24. [24]Santamaria L. Martinez-Onsurbe P. Paniagua R. Nistal M. Laminin, type iv collagen, and fibronectin in normal and cryptorchid human testes. An immunohistochemical study Int. J. Androl. 1990 13 135 146 10.1111/j.1365-2605.1990.tb00970.x 1971616 · DOI
  25. [25]Hussein K.H. Park K.M. Kang K.S. Woo H.M. Biocompatibility evaluation of tissue-engineered decellularized scaffolds for biomedical application Mater. Sci. Eng. C Mater. Biol. Appl. 2016 67 766 778 10.1016/j.msec.2016.05.068 27287176 · DOI · Paper page
  26. [26]Crapo P.M. Gilbert T.W. Badylak S.F. An overview of tissue and whole organ decellularization processes Biomaterials 2011 32 3233 3243 10.1016/j.biomaterials.2011.01.057 21296410 PMC3084613 · DOI · Paper page
  27. [27]Brown B.N. Badylak S.F. Extracellular matrix as an inductive scaffold for functional tissue reconstruction Transl. Res. 2014 163 268 285 10.1016/j.trsl.2013.11.003 24291155 PMC4203714 · DOI
  28. [28]Baert Y. Stukenborg J.B. Landreh M. De Kock J. Jornvall H. Soder O. Goossens E. Derivation and characterization of a cytocompatible scaffold from human testis Hum. Reprod. 2015 30 256 267 10.1093/humrep/deu330 25505010 · DOI · Paper page
  29. [29]Baert Y. De Kock J. Alves-Lopes J.P. Soder O. Stukenborg J.B. Goossens E. Primary human testicular cells self-organize into organoids with testicular properties Stem Cell Rep. 2016 8 30 38 10.1016/j.stemcr.2016.11.012 28017656 PMC5233407 · DOI
  30. [31]Liao J. Joyce E.M. Sacks M.S. Effects of decellularization on the mechanical and structural properties of the porcine aortic valve leaflet Biomaterials 2008 29 1065 1074 10.1016/j.biomaterials.2007.11.007 18096223 PMC2253688 · DOI
  31. [34]Hadley M.A. Weeks B.S. Kleinman H.K. Dym M. Laminin promotes formation of cord-like structures by sertoli cells in vitro Dev. Biol. 1990 140 318 327 10.1016/0012-1606(90)90082-T 2373256 · DOI
  32. [35]van der Wee K. Hofmann M.C. An in vitro tubule assay identifies hgf as a morphogen for the formation of seminiferous tubules in the postnatal mouse testis Exp. Cell Res. 1999 252 175 185 10.1006/excr.1999.4630 10502410 · DOI
  33. [36]Chui K. Trivedi A. Cheng C.Y. Cherbavaz D.B. Dazin P.F. Huynh A.L. Mitchell J.B. Rabinovich G.A. Noble-Haeusslein L.J. John C.M. Characterization and functionality of proliferative human sertoli cells Cell Trans. 2011 20 619 635 10.3727/096368910X536563 21054948 PMC4096632 · DOI
  34. [37]Ren H. Shi X. Tao L. Xiao J. Han B. Zhang Y. Yuan X. Ding Y. Evaluation of two decellularization methods in the development of a whole-organ decellularized rat liver scaffold Liver Int. 2013 33 448 458 10.1111/liv.12088 23301992 · DOI · Paper page
  35. [38]Oberwallner B. Brodarac A. Choi Y.H. Saric T. Anic P. Morawietz L. Stamm C. Preparation of cardiac extracellular matrix scaffolds by decellularization of human myocardium J. Biomed. Mater. Res. A 2014 102 3263 3272 10.1002/jbm.a.35000 24142588 · DOI
  36. [39]Svingen T. Koopman P. Building the mammalian testis: Origins, differentiation, and assembly of the component cell populations Genes Dev. 2013 27 2409 2426 10.1101/gad.228080.113 24240231 PMC3841730 · DOI
  37. [40]Enders G.C. Henson J.H. Millette C.F. Sertoli cell binding to isolated testicular basement membrane J. Cell Biol. 1986 103 1109 1119 10.1083/jcb.103.3.1109 3528169 PMC2114291 · DOI
  38. [41]Mohiuddin M.M. Singh A.K. Corcoran P.C. Hoyt R.F. Thomas M.L. 3rd Ayares D. Horvath K.A. Genetically engineered pigs and target-specific immunomodulation provide significant graft survival and hope for clinical cardiac xenotransplantation J. Thorac. Cardiovasc. Surg. 2014 148 1106 1113 10.1016/j.jtcvs.2014.06.002 24998698 PMC4135017 · DOI
  39. [42]Platz J. Bonenfant N.R. Uhl F.E. Coffey A.L. McKnight T. Parsons C. Sokocevic D. Borg Z.D. Lam Y.W. Deng B. Comparative decellularization and recellularization of wild-type and alpha 1,3 galactosyltransferase knockout pig lungs: A model for ex vivo xenogeneic lung bioengineering and transplantation Tissue Eng. Part C Methods 2016 22 725 739 10.1089/ten.tec.2016.0109 27310581 PMC4991572 · DOI
  40. [43]Schlatt S. de Kretser D.M. Loveland K.L. Discriminative analysis of rat sertoli and peritubular cells and their proliferation in vitro: Evidence for follicle-stimulating hormone-mediated contact inhibition of sertoli cell mitosis Biol. Reprod. 1996 55 227 235 10.1095/biolreprod55.2.227 8828824 · DOI
  41. [44]Feng L.X. Ravindranath N. Dym M. Stem cell factor/c-kit up-regulates cyclin d3 and promotes cell cycle progression via the phosphoinositide 3-kinase/p70 s6 kinase pathway in spermatogonia J. Biol. Chem. 2000 275 25572 25576 10.1074/jbc.M002218200 10849422 · DOI
  42. [45]Pendergraft S.S. Sadri-Ardekani H. Atala A. Bishop C.E. Three-dimensional testicular organoid: A novel tool for the study of human spermatogenesis and gonadotoxicity in vitro Biol. Reprod. 2017 96 720 732 10.1095/biolreprod.116.143446 28339648 · DOI
  43. [46]Alves-Lopes J.P. Soder O. Stukenborg J.B. Testicular organoid generation by a novel in vitro three-layer gradient system Biomaterials 2017 130 76 89 10.1016/j.biomaterials.2017.03.025 28364632 · DOI

Authors
Vermeulen, M., Del Vento, F., De Michele, F., Poels, J., & Wyns, C.
Affiliations
  • UCLouvain (BE)
  • Gynecology-Andrology Research Unit, Institut de Recherche Expérimentale et Clinique, Medical School, Université Catholique de Louvain, 1200 Brussels, Belgium
  • Cliniques Universitaires Saint-Luc (BE)
Corresponding author
Wyns, Christine
ORCID
Venue
International Journal of Molecular Sciences
Publisher
MDPI AG
Published
2018-01-12
Volume
19
Issue
1
Pages
227
ISSN
1422-0067 (ISSN-L)
DOI
10.3390/ijms19010227
PMC
PMC5796176
OpenAlex
W2782965431
Cited by
72
References
47
Citations by year
2026: 4 · 2025: 7 · 2024: 4 · 2023: 9 · 2022: 10 · 2021: 15 · 2020: 10 · 2019: 8 · 2018: 5
FWCI
9.88
Type
journal article
Language
en
Open access
Licence
Locations
  • International Journal of Molecular Sciences (publishedVersion, cc-by)
  • Europe PMC (PubMed Central) (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • Digital Access to Libraries (submittedVersion, other-oa)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
Funders
Fonds De La Recherche Scientifique - FNRS (7451016F)
Topics
  • Health Sciences › Medicine › Surgery › Tissue Engineering and Regenerative Medicine
  • Physical Sciences › Engineering › Biomedical Engineering › 3D Printing in Biomedical Research
  • Health Sciences › Medicine › Reproductive Medicine › Sperm and Testicular Function
Keywords
decellularization, Sertoli cells, fertility preservation, cancer patients, extracellular matrix preservation, glycosaminoglycans, stem cell factor, Sertoli cell proliferation, xenotransplantation
MeSH terms
Adult, Animals, Biocompatible Materials, Cell Adhesion, DNA, Extracellular Matrix, Glycosaminoglycans, Humans, Male, Sertoli Cells, Testis, Sus scrofa, Cell Proliferation, Tissue Scaffolds

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

Are you an author, or do you see a problem with this page? Request a review