Data_Sheet_1_A Natural Xenogeneic Endometrial Extracellular Matrix Hydrogel Toward Improving Current Human in vitro Models and Future in vivo Applications.zip

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.

脱细胞技术（decellularization techniques）可用于制备生物相容性细胞外基质（extracellular matrix）水凝胶，为体外（in vitro）细胞培养与体内（in vivo）组织再生提供组织特异性微环境。本研究从猪源脱细胞子宫中提取子宫内膜组织，用以制备子宫内膜细胞外基质（endometrial extracellular matrix，简称EndoECM）水凝胶。在完成脱细胞与去污剂清除流程后，我们对EndoECM的理化特性展开了研究，涵盖凝胶动力学、超微结构以及蛋白质组学特征（proteomic profile）。其基质组（matrisome）显示保留了结构组分与组织特异性成分，且免疫反应性分子含量极低。EndoECM可支持人子宫内膜细胞在二维与三维培养体系中的体外培养，相较于胶原蛋白与基质胶（Matrigel），更能促进子宫内膜干细胞的增殖。此外，我们构建了源自不同来源的上皮细胞与基质细胞的三维子宫内膜样共培养体系，该共培养体系可维持细胞活性，并呈现出显著的组织重塑能力。最后，在一项初步研究中，我们将EndoECM皮下注射至免疫健全小鼠体内，以检测其潜在的低免疫原性反应。仿生子宫内膜微环境为生殖技术与子宫内膜修复提供了全新策略，本研究结果证实EndoECM在子宫内膜体外培养以及子宫内膜病变治疗方面具有应用潜力。