title: Rat parotid gland cell differentiation in three-dimensional culture.
abstract: The use of polarized salivary gland cell monolayers has contributed to our understanding of salivary gland physiology. However, these cell models are not representative of glandular epithelium in vivo, and, therefore, are not ideal for investigating salivary epithelial functions. The current study has developed a three-dimensional (3D) cell culture model for rat Par-C10 parotid gland cells that forms differentiated acinar-like spheres on Matrigel. These 3D Par-C10 acinar-like spheres display characteristics similar to differentiated acini in salivary glands, including cell polarization, tight junction (TJ) formation required to maintain transepithelial potential difference, basolateral expression of aquaporin-3 and Na+/K+/2Cl- cotransporter-1, and responsiveness to the muscarinic receptor agonist carbachol that is decreased by the anion channel blocker diphenylamine-2-carboxylic acid or chloride replacement with gluconate. Incubation of the spheres in the hypertonic medium increased the expression level of the water channel aquaporin-5. Further, the proinflammatory cytokines tumor necrosis factor-alpha and interferon-gamma induced alterations in TJ integrity in the acinar-like spheres without affecting individual cell viability, suggesting that cytokines may affect salivary gland function by altering TJ integrity. Thus, 3D Par-C10 acinar-like spheres represent a novel in vitro model to study physiological and pathophysiological functions of differentiated acini.
authors.affiliation: Department of Oral Biology, School of Dental Medicine, University at Buffalo, The State University of New York, Buffalo, New York 14214-3092, USA. olgabake@buffalo.edu
authors.forename: Olga J
authors.initials: OJ
authors.lastname: Baker
authors.forename: David J
authors.initials: DJ
authors.lastname: Schulz
authors.forename: Jean M
authors.initials: JM
authors.lastname: Camden
authors.forename: Zhongji
authors.initials: Z
authors.lastname: Liao
authors.forename: Troy S
authors.initials: TS
authors.lastname: Peterson
authors.forename: Cheikh I
authors.initials: CI
authors.lastname: Seye
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Petris
authors.forename: Gary A
authors.initials: GA
authors.lastname: Weisman
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002217:Carbachol / Q000494:pharmacology; D002454:Cell Differentiation; D016764:Cell Polarity; D002478:Cells, Cultured; D007371:Interferon-gamma / Q000494:pharmacology; D018613:Microscopy, Confocal; D010306:Parotid Gland / Q000166:cytology*; D051381:Rats; D019108:Tight Junctions / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology; D062367:ortho-Aminobenzoates / Q000494:pharmacology
chemical_list: D014409:Tumor Necrosis Factor-alpha; D062367:ortho-Aminobenzoates; D007371:Interferon-gamma; D002217:Carbachol; C001209:fenamic acid