Employing a GelMA/Decellularized ECM Biomaterial to Enhance In Vitro Modeling Approaches in Reproductive Biosciences.

Climate change poses a significant threat to global health, with pregnant women and their offspring being particularly vulnerable to heat stress. This rising concern has created an urgent need for advanced in vitro models to study how climate change-associated heat stress affects maternal and offspring health, focusing on how in utero heat stress impacts life course health. The tissue that mediates this maternal stress to the offspring is the endometrium - the innermost lining of the uterus and is comprised of a larger number of phenotypically diverse cells. This work develops a biomaterial-based platform to better mimic the endometrium. We synthesized optimized methacrylated gelatin (GelMA) hydrogel for bovine endometrial stromal cell (ESC) culture. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays demonstrated the suitability of 10% (w/v) GelMA hydrogels for bovine ESC culture. To better mimic the native endometrial microenvironment, we decellularized extracellular matrix (dECM) from porcine endometrium and developed a novel bicomponent system combining GelMA with endometrial dECM. Physical characterization of the hydrogels indicated that dECM incorporation significantly affected hydrogel properties. It decreased the transparency of the scaffolds, creating a denser polymeric network and reducing the degradation rate. Additionally, the presence of dECM in the structure, even at a low concentration of 4 mg/mL, significantly improved the biocompatibility of the scaffolds, as evidenced by greater bovine ESC viability within the GelMA/dECM hydrogels compared to GelMA hydrogels alone.Clinical Relevance-The developed hydrogel could be used for the development of endometrial organoids and the co-culture of endometrial cells within an endometrium-on-a-chip model to investigate heat stress effects on the endometrium.