Enhancing Insight into Human Endometrial Biology through advanced 3D Cellular Models

Embryo implantation has been defined as the "black box" of human reproduction. Most of the knowledge on mechanisms underlying this process derives from animal models, which cannot always be translated to humans. Therefore, new technologies such as the 3D cell models are considered a new step to foster the study of human endometrial biology and an advanced tool for studying endometrial-associated diseases and understanding the complex mechanisms surrounding endometrium-embryo crosstalk. The first aim of this thesis was to investigate a noninvasive model of the endometrium using human primary cells in static conditions. The menstrual flow was used as a source to establish a reliable and complete human 3D model. In fact, from human menstrual flow, is possible to isolate both stromal and epithelial cells to generate the 3D model. Subsequently, this model was characterized and stimulated to mimic different phases of the menstrual cycle and resulted in being as responsive as the biopsy counterpart. The second aim of this thesis is to develop a 3D model of the human endometrium using Microfluidic devices to better recapitulate the physiological changes occurring during the menstrual cycle. The decidualization process has been a focal point and the two main cell populations of the endometrium have been considered: the endometrial epithelial cell line EM42, and the immortalized endometrial stromal cell line THESC. The survival and reactivity of the model have been tested as well, aware of the lack of important cellular interactions necessary to fully mimic the endometrial environment in toto. The model was also enriched with the addition of HUtMECs, endothelial cells deeply involved in the vascularization process. In the future other cell types should be included in this model to a micro-physiological-system (MPF), for example, the immune cells.