Human Uterine Epithelial Cells: Influence of culture conditions and stromal cells on epithelial cell transepithelial cell resistance

Abstract

The purpose of this study was to determine the effects of extracellular matrices, media formulations, menstrual cycle and menopausal status, and culture with stromal cells on the growth and transepithelial resistance of human uterine epithelial cells in cell chambers. Human uterine epithelial cells were obtained following hysterectomies from pre- and post-menopausal women, and were cultured in the apical chamber of extracellular-matrix coated cell chambers. Matrigel, Reduced Growth Factor-Matrigel, and Human Extracellular Matrix were sufficient to support cell growth to confluence, tight junction formation and high values of transepithelial resistance. DMEM-F12 complete medium was superior to Leibowitz complete medium for obtaining reproducible cell growth. Uterine epithelial cells from women in the secretory and proliferative stages of the menstrual cycle and in post-menopause grew equally well. Growing epithelial sheets in the apical chamber above stromal cells in the basolateral chamber inhibited cell growth and the development of transepithelial resistance. Similarly, placing confluent epithelial cells over stromal cells from the same patient resulted in a decrease in transepithelial resistance. Conversely, epithelial cells initially cultured with stromal cells in the basolateral compartment developed high transepithelial resistance when stromal cells were removed. These results identify the optimal conditions for human uterine epithelial cell growth to confluence and polarization. Further, these findings demonstrate that epitheliahstromal interactions influence epithelial cell integrity as measured by transepithelial resistance.p