The goal of this project was to develop in vitro models of the female genital tract epithelia to study bacterial colonisation. The vaginal microbiota has been widely described but few models are available to study how it impacts the vaginal epithelium. It is only recently that the existence of an endometrial microbiota was discovered so its composition and importance remain elusive. L. crispatus and G. vaginalis have both been described in the vaginal microbiota as being associated with a healthy vaginal state and with bacterial vaginosis respectively. Here, we describe the establishment of an endometrial in vitro model with -estradiol that supports bacterial colonisation by L. crispatus and G. vaginalis. Assessment of cell viability did not show major differences between the two strains. We also discuss an isolation protocol for primary vaginal epithelial cells to reproduce the in vitro model and procedure of bacterial colonisation. Providing simple, accessible, and reproductible in vitro models of the FGT is essential to study genital microbiota interactions and how probiotic or other drugs might affect it.