title: Coculturing human endometrial epithelial cells and stromal fibroblasts alters cell-specific gene expression and cytokine production.
abstract: OBJECTIVE
  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.
  
  
  METHODS
  In vitro study.
  
  
  METHODS
  University research laboratory.
  
  
  METHODS
  Endometrial biopsies were obtained from premenopausal women.
  
  
  METHODS
  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.
  
  
  METHODS
  Cell-specific global gene expression profiling and analysis of secreted cytokines in eEC/eSF cocultures and respective monocultures.
  
  
  RESULTS
  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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Department of Obstetrics, Gynecology and Reproductive Sciences, University of California, Center for Reproductive Sciences, San Francisco, California.
authors.forename: Joseph C
authors.initials: JC
authors.lastname: Chen
authors.forename: David W
authors.initials: DW
authors.lastname: Erikson
authors.forename: Terhi T
authors.initials: TT
authors.lastname: Piltonen
authors.forename: Michelle R
authors.initials: MR
authors.lastname: Meyer
authors.forename: Fatima
authors.initials: F
authors.lastname: Barragan
authors.forename: Ramsey H
authors.initials: RH
authors.lastname: McIntire
authors.forename: John S
authors.initials: JS
authors.lastname: Tamaresis
authors.forename: Kim Chi
authors.initials: KC
authors.lastname: Vo
authors.forename: Linda C
authors.initials: LC
authors.lastname: Giudice
authors.forename: Juan C
authors.initials: JC
authors.lastname: Irwin
mesh_terms: D000328:Adult; D002450:Cell Communication; D016764:Cell Polarity; D002469:Cell Separation / Q000379:methods; D048430:Cell Shape; D002478:Cells, Cultured; D018920:Coculture Techniques; D017077:Culture Media, Conditioned / Q000378:metabolism; D016207:Cytokines / Q000235:genetics / Q000378:metabolism*; D004717:Endometrium / Q000166:cytology / Q000276:immunology / Q000378:metabolism*; D004847:Epithelial Cells / Q000276:immunology / Q000378:metabolism*; D005260:Female; D005347:Fibroblasts / Q000276:immunology / Q000378:metabolism*; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D020869:Gene Expression Profiling / Q000379:methods; D005786:Gene Expression Regulation; D006801:Humans; D008875:Middle Aged; D020411:Oligonucleotide Array Sequence Analysis; D017154:Stromal Cells / Q000276:immunology / Q000378:metabolism*; D013997:Time Factors
chemical_list: D017077:Culture Media, Conditioned; D016207:Cytokines
keywords: Endometrium; coculture; cytokines; microarray; polarized epithelium; stroma