Endocervical crypts secrete cervical mucus whose biochemical, biophysical and functional properties change depending on the phase of the menstrual cycle, so it is required to use an appropriate experimental model to investigate the secretory activity of the endocervix. The development of 3D cell cultures has enabled the cultivation of highly differentiated and functional endocervical cells with clearly defined polarity, intercellular junctions, and mucin secretion ability. To achieve a successful 3D culture, it is necessary to optimize specific conditions. In two consecutive cultivations, the optimal experimental settings for culturing the human epithelial endocervical cell line End1 / E6E7 in 3D culture were examined using transwell insert that allows cells growth at the "medium - air" interface. Different concentrations and times of exposure to extracellular matrix were examined, as well as different exposure time to ß-estradiol and incubation time at which cells achieve complete differentiation and functionality. Cell growth, differentiation and functionality were assessed by cytological, cytochemical, immunocytochemical methods and electron microscopy. With this thesis optimal conditions for growth, differentiation and functionalization of endocervical cells in 3D culture were established. The first signs of differentiation and functionalization appeared on the sixth day of cultivation, and full functionality was achieved by the 18th day. The uniformity of mucin production in the cells enabled the application of the established 3D culture in the experiment of cell exposure to ß-estradiol. Exposed cells improved the formation of tight and adherent junctions and reduced the amount of mucin within the cells.