title: Cytoarchitecture modifications of the human uterine endocervical mucosa in long-term three-dimensional organotypic culture.
abstract: We assayed the effects of phenol red (pr), estrogen (Es), and progesterone (Pg) in three-dimensional organotypic cultures of human uterine endocervix. Small intact fragments deposited on sponges submerged in DMEM with 10% fetal bovine serum were cultured in three different conditions: with pr (DMEM(pr+)), without pr (DMEM(pr-)), and without pr but with the addition of physiological concentrations of Es and Pg [DMEM(pr-)(Es+Pg)]. Cell viability and cellular responses were assayed after 4, 10, and 21 days using immunohistochemistry for the expression and distribution of the following markers: mucins and mucopolysaccharides (PAS staining), pan-cytokeratins (AE1/AE3), CK19, p63, Ki-67, vimentin, estrogen receptor-alpha (ER-alpha), and progesterone receptor (PR). The fragments cultivated in DMEM(pr+) showed a cuboidal, poorly differentiated epithelial phenotype and signs of stroma degeneration. In DMEM(pr-), both tissue architecture and cellular heterogeneity were much better preserved: epithelial cells showed a more columnar shape, and stroma was very well conserved, maintaining cell density. The addition of Es and Pg further improved the histology and physiology of the fragments: in DMEM(pr-)(Es+Pg), epithelial cells retained a columnar shape, secreted mucins, and formed areas of squamous hyperplasia.
authors.affiliation: Immunobiology and Cell Differentiation Unit, Institute of Biomedical Technologies, CNR, Via G. Moruzzi 1, 56100 Pisa, Italy.
authors.forename: Monica
authors.initials: M
authors.lastname: Michelini
authors.forename: Alfredo
authors.initials: A
authors.lastname: Rosellini
authors.forename: Vaclav
authors.initials: V
authors.lastname: Mandys
authors.forename: Tommaso
authors.initials: T
authors.lastname: Simoncini
authors.forename: Roberto P
authors.initials: RP
authors.lastname: Revoltella
mesh_terms: D015415:Biomarkers / Q000378:metabolism; D002452:Cell Count; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002584:Cervix Uteri / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D004967:Estrogens / Q000494:pharmacology; D005260:Female; D006801:Humans; D007124:Immunoenzyme Techniques; D009077:Mucins / Q000378:metabolism; D009092:Mucous Membrane / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D009924:Organ Culture Techniques / Q000379:methods*; D010637:Phenolsulfonphthalein / Q000494:pharmacology; D017697:Premenopause; D011374:Progesterone / Q000494:pharmacology
chemical_list: D015415:Biomarkers; D004967:Estrogens; D009077:Mucins; D011374:Progesterone; D010637:Phenolsulfonphthalein