Fig. 3

Culture of endometrial epithelial cells with endometrial stromal cells. a, b Microscopic appearance of endometrial stromal cells cultured in conventional medium (DMEM) (a) and epithelium-specific medium (ESTEM-HE medium) (b). Black bar is 500 μm. c Growth curves of endometrial stromal cells cultured in conventional and epithelium-specific medium. Error bar indicates SEM. An asterisk means P < 0.05. d Microscopic appearance of endometrial epithelial cells with endometrial stromal cells in serial passage. Black bar is 500 μm. e–g Cumulative area of colonies (e), colony formation (number) (f), and area of colonies (g) of endometrial epithelial cells in serial passage with endometrial stromal cells. Error bar indicates SEM. An asterisk means P < 0.05. h Immunocytochemical staining for endometrial epithelial cells and endometrial stromal cells at passage 4. Endometrial epithelial cells (surrounded with white dotted lines) continued to express pan-cytokeratin, but not vimentin, at passage 4. Endometrial stromal cells were positive for vimentin. Nuclei were stained with DAPI. Yellow bar is 500 μm. Each experiment was done in triplicate. Abbreviation: DMEM, Dulbecco’s modified Eagle’s medium; SEM, standard error of the mean