According to the World Health Organization, endometriosis is a disease affecting 10% of women worldwide. Endometriosis symptoms impacts very negatively the patient’s quality of life and, despite its high prevalence, diagnostic methods and treatments are very limited. In order to improve them, models are needed to help research. In fact, it exists no satisfying in vivo or in vitro model that integrates all the components of human endometriosis disease. The MOON Project aims to develop an in vitro model that respond to this need. In this work, we aim to select endometrial cell lines, characterize them and design co-culture models usable in the project. Ishikawa and ESS-1 cell lines were chosen as epithelial and stromal endometrial cell line respectively. Ishikawa cells characterization shows that this epithelial cell line doesn’t form impermeable epithelium, is able to survive on dynamic flow but with some mortality and is able to invade other tissue. Moreover, their growth doesn’t seem impacted by estradiol and progesterone treatment. We observed, nevertheless, Progesterone Receptor induction by estradiol treatment. Concerning co-culture models, we assessed that all cells line selected can grow in the same culture medium. Moreover, we chose different markers to differentiate them and tested it by immunofluorescence. Finally, we managed to create 3D model constructs based on a collagen matrix.