Abstract
Decellularization techniques support the creation of biocompatible extracellular matrix hydrogels, providing tissue-specific environments for both in vitro cell culture and in vivo tissue regeneration. We obtained endometrium derived from porcine decellularized uteri to create endometrial extracellular matrix (EndoECM) hydrogels. After decellularization and detergent removal, we investigated the physicochemical features of the EndoECM, including gelation kinetics, ultrastructure, and proteomic profile. The matrisome showed conservation of structural and tissue-specific components with low amounts of immunoreactive molecules. EndoECM supported in vitro culture of human endometrial cells in two- and three-dimensional conditions and improved proliferation of endometrial stem cells with respect to collagen and Matrigel. Further, we developed a three-dimensional endometrium-like co-culture system of epithelial and stromal cells from different origins. Endometrial co-cultures remained viable and showed significant remodeling. Finally, EndoECM was injected subcutaneously in immunocompetent mice in a preliminary study to test a possible hypoimmunogenic reaction. Biomimetic endometrial milieus offer new strategies in reproductive techniques and endometrial repair and our findings demonstrate that EndoECM has potential for in vitro endometrial culture and as treatment for endometrial pathologies.
Highlights
The extracellular matrix (ECM) is a complex mixture of fibrous proteins and acts as a physical substrate for cell adhesion, structure, and mechanical stimulation
Decellularization efficiency was verified by Hematoxylin and Eosin (H&E) (Figures 3B1,2) and Masson’s Trichrome (MT) staining (Figures 3B3,4), which showed complete depletion of cellular material, while DAPI staining showed a total removal of cell nuclei (Figures 3B5,6)
Blue coloration in MT staining in DC tissues indicates collagen conservation, a principal ECM component
Summary
The extracellular matrix (ECM) is a complex mixture of fibrous proteins and acts as a physical substrate for cell adhesion, structure, and mechanical stimulation It provides biochemical cues for tissue homeostasis, angiogenesis, immune response, and tissue repair (Frantz et al, 2010; Rozario and DeSimone, 2010; Evangelatov and Pankov, 2013; Theocharis et al, 2016; Saldin et al, 2017). Three-dimensional models used to recreate the endometrium in vitro incorporate ECMderived matrices such as collagen I and basement membrane extract (Schutte and Taylor, 2012; Asmani et al, 2013; Bayat et al, 2016; Fayazi et al, 2017) These standardized matrices do not have the biochemical complexity of the natural endometrial ECM and are not suitable for clinical application because they originate from a tumorigenic cell line
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