Coculturing human endometrial epithelial cells and stromal fibroblasts alters cell-specific gene expression and cytokine production

• Published in: Fertility and sterility Vol. 100; no. 4; pp. 1132 - 1143
• Main Authors: Chen, Joseph C., Erikson, David W., Piltonen, Terhi T., Meyer, Michelle R., Barragan, Fatima, McIntire, Ramsey H., Tamaresis, John S., Vo, Kim Chi, Giudice, Linda C., Irwin, Juan C.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.10.2013
• Subjects: Adult; Cell Communication; Cell Polarity; Cell Separation - methods; Cell Shape; Cells, Cultured; coculture; Coculture Techniques; Culture Media, Conditioned - metabolism; cytokines; Cytokines - genetics; Cytokines - metabolism; Endometrium; Endometrium - cytology; Endometrium - immunology; Endometrium - metabolism; epithelial cells; Epithelial Cells - immunology; Epithelial Cells - metabolism; epithelium; Female; fibroblasts; Fibroblasts - immunology; Fibroblasts - metabolism; Flow Cytometry; Fluorescent Antibody Technique; gene expression; Gene Expression Profiling - methods; Gene Expression Regulation; Humans; in vitro studies; Internal Medicine; microarray; Middle Aged; Obstetrics and Gynecology; Oligonucleotide Array Sequence Analysis; patients; polarized epithelium; premenopause; proteins; secretion; stroma; stromal cells; Stromal Cells - immunology; Stromal Cells - metabolism; tight junctions; Time Factors; transcriptome; university research; women; polarized epithelium; microarray; cytokines; coculture; Endometrium; stroma
• ISSN: 0015-0282; 1556-5653; 1556-5653
• DOI: 10.1016/j.fertnstert.2013.06.007

To determine the effects of coculturing endometrial epithelial cells (eEC) with paired endometrial stromal fibroblasts (eSF) on cell-specific gene expression and cytokine secretion patterns. In vitro study. University research laboratory. Endometrial biopsies were obtained from premenopausal women. Polarized eEC and subject-paired eSF were cultured for 12.5 hours alone (monoculture) or combined in a two-chamber coculture system without cell-cell contact. Cells and conditioned media were analyzed for global gene expression and cytokine secretion, respectively. Purified, endometrial tissue-derived eEC and eSF isolated by fluorescent activated cell sorting (FACS) were used as noncultured controls. Cell-specific global gene expression profiling and analysis of secreted cytokines in eEC/eSF cocultures and respective monocultures. Transepithelial resistance, diffusible tracer exclusion, expression of tight junction proteins, and apical/basolateral vectorial secretion confirmed eEC structural and functional polarization. Distinct transcriptomes of eEC and eSF were consistent with their respective lineages and their endometrial origin. Coculture of eEC with eSF resulted in altered cell-specific gene expression and cytokine secretion. This coculture model provides evidence that interactions between endometrial functionally polarized epithelium and stromal fibroblasts affect cell-specific gene expression and cytokine secretion underscoring their relevance when modeling endometrium in vitro.