• A protocol for effective decellularization of human endometrial fragments for clinical applications
  • Zinat Sargazi,1,* saeed zavareh,2 mojdeh salehnia,3
    1. department of basic medical sciences, neyshabur university of medical sciences, neyshabur, iran
    3. Professor of Histology and Embryology


  • Introduction: The objective of this study was to evaluate and compare the effectiveness of various decellularization protocols when applied to human endometrial fragments.
  • Methods: decellularization process involved a combination of freeze-thaw cycles, Triton X-100 treatment, and four different concentrations of sodium dodecyl sulfate (SDS; 0.1%, 0.5%, 1%, and 1.5%) applied for two distinct exposure durations (24 and 72 hours). Following analysis of tissue morphology and DNA content, The tissue group with the best morphology and lowest DNA content was chosen for further analysis. The nucleus was visualized using Acridine orange staining, and the extracellular matrix (ECM) was examined using Masson's trichrome, Alcian blue, and periodic acid-Schiff staining. Raman spectroscopy was used to quantify the levels of collagen types I and IV, fibronectin, glycosaminoglycans (GAGs), and elastin in the tissues. The ultrastructure and porosity of the decellularized scaffold were examined using scanning electron microscopy (SEM), and its cytotoxicity was assessed with an MTT assay.
  • Results: The treated group with 1% SDS for 72 h showed the morphology similar to native control in having the minimum level of DNA and well preserved ECM. Raman spectroscopy analysis revealed no significant difference in the amounts of collagen types I and IV, GAG, and fibronectin between the decellularized scaffold and the native group but the elastin protein level was significantly decreased (P < 0.001). SEM micrographs also showed a porous and fiber rich ECM in decellularized sample similar to the native control.
  • Conclusion: This combined protocol for decellularization of human endometrial tissue is effective and it could be suitable for recellularization and clinical applications in the future.
  • Keywords: Decellularization; Human endometrial tissue; Raman spectroscopy; Sodium dodecyl sulfate