Specific biological properties of ovarian cancer cells can be modeled and studied using in vitro experiments. Any experimental setting can closely reflect some aspects of the native conditions; however, parameters that differ from in vivo aspects must be considered. Familiarity with existing and well-established, as well as new, cell culture techniques provides a basis for correct experimental design and production of reliable scientific results. This chapter presents a short comparative review of the techniques used for cell culture establishment and maintenance of ovarian cancer cells, as well as laboratory methods used to characterize malignant features of these cells, including the epithelial–mesechymal transition, cell motility and invasiveness, resistance to detachment-induced apoptosis, and stem cell content.