title: Matrigel improves functional properties of primary human salivary gland cells.
abstract: Currently, there is no effective treatment available to patients with irreversible loss of functional salivary acini caused by Sjogren's syndrome or after radiotherapy for head and neck cancer. A tissue-engineered artificial salivary gland would help these patients. The graft cells for this device must establish tight junctions in addition to being of fluid-secretory nature. This study analyzed a graft source from human salivary glands (huSG) cultured on Matrigel. Cells were obtained from parotid and submandibular glands, expanded in vitro, and then plated on either Matrigel-coated (2 mg/mL) or uncoated culture dish. Immunohistochemistry, transmission electron microscopy, quantitative real-time-polymerase chain reaction, Western blot, and transepithelial electrical resistance were employed. On Matrigel, huSG cells adopted an acinar phenotype by forming three-dimensional acinar-like units (within 24 h of plating) as well as a monolayer of cells. On uncoated surfaces (plastic), huSG cells only formed monolayers of ductal cells. Both types of culture conditions allowed huSG cells to express tight junction proteins (claudin-1, -2, -3, -4; occludin; JAM-A; and ZO-1) and adequate transepithelial electrical resistance. Importantly, 99% of huSG cells on Matrigel expressed α-amylase and the water channel protein Aquaporin-5, as compared to <5% of huSG cells on plastic. Transmission electron microscopy confirmed an acinar phenotype with many secretory granules. Matrigel increased the secretion of α-amylase two to five folds into the media, downregulated certain salivary genes, and regulated the translation of acinar proteins. This three-dimensional in vitro serum-free cell culture method allows the organization and differentiation of huSG cells into salivary cells with an acinar phenotype.
authors.affiliation: Faculty of Dentistry, McGill University, Montreal, Canada.
authors.forename: Ola M
authors.initials: OM
authors.lastname: Maria
authors.forename: Anthony
authors.initials: A
authors.lastname: Zeitouni
authors.forename: Olga
authors.initials: O
authors.lastname: Gologan
authors.forename: Simon D
authors.initials: SD
authors.lastname: Tran
mesh_terms: D000943:Antigens, Differentiation / Q000096:biosynthesis; D001672:Biocompatible Materials / Q000737:chemistry*; D002478:Cells, Cultured; D003094:Collagen / Q000737:chemistry*; D004338:Drug Combinations; D005260:Female; D006801:Humans; D007797:Laminin / Q000737:chemistry*; D008297:Male; D011509:Proteoglycans / Q000737:chemistry*; D012859:Sjogren's Syndrome / Q000378:metabolism / Q000628:therapy; D013363:Submandibular Gland / Q000166:cytology* / Q000378:metabolism*; D023822:Tissue Engineering / Q000379:methods*
chemical_list: D000943:Antigens, Differentiation; D001672:Biocompatible Materials; D004338:Drug Combinations; D007797:Laminin; D011509:Proteoglycans; C059485:matrigel; D003094:Collagen