title: Differentiation of primary human submandibular gland cells cultured on basement membrane extract.
abstract: There is no effective treatment for the loss of functional salivary tissue after irradiation for head and neck cancer or the autoimmune disease Sjögren's syndrome. One possible approach is the regeneration of salivary glands from stem cells. The present study aimed to investigate whether small pieces of human submandiblar gland tissue contain elements necessary for the reconstruction of salivary rudiments in vitro via acinar and ductal cell differentiation. Primary submandibular gland (primary total human salivary gland; PTHSG) cells were isolated from human tissue and cultured in vitro using a new method in which single cells form an expanding epithelial monolayer on plastic substrates. Differentiation, morphology, number, and organization of these cells were then followed on basement membrane extract (BME) using RNA quantitation (amylase, claudin-1 (CLN1), CLN3, kallikrein, vimentin), immunohistochemistry (amylase and occludin), viability assay, and videomicroscopy. On the surface of BME, PTHSG cells formed acinotubular structures within 24 h, did not proliferate, and stained for amylase. In cultures derived from half of the donors, the acinar markers amylase and CLN3 were upregulated. The PTHSG culture model suggests that human salivary gland may be capable of regeneration via reorganization and differentiation and that basement membrane components play a crucial role in the morphological and functional differentiation of salivary cells.
authors: Department of Oral Biology, Semmelweis University and Hungarian Academy of Sciences, Budapest, Hungary.
authors: Vanda
authors: V
authors: Szlávik
authors: Bálint
authors: B
authors: Szabó
authors: Tamás
authors: T
authors: Vicsek
authors: József
authors: J
authors: Barabás
authors: Sándor
authors: S
authors: Bogdán
authors: Veronika
authors: V
authors: Gresz
authors: Gábor
authors: G
authors: Varga
authors: Brian
authors: B
authors: O'Connell
authors: János
authors: J
authors: Vág
mesh_terms: D000681:Amylases / Q000235:genetics / Q000378:metabolism; D001485:Basement Membrane / Q000378:metabolism*; D002454:Cell Differentiation / Q000235:genetics / Q000502:physiology*; D049109:Cell Proliferation; D002470:Cell Survival; D062445:Claudin-1; D006801:Humans; D007150:Immunohistochemistry; D007610:Kallikreins / Q000235:genetics / Q000378:metabolism; D008562:Membrane Glycoproteins / Q000235:genetics / Q000378:metabolism; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism; D018832:Molecular Chaperones / Q000235:genetics / Q000378:metabolism; D062793:Occludin; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013363:Submandibular Gland / Q000166:cytology* / Q000378:metabolism; D014746:Vimentin / Q000235:genetics / Q000378:metabolism
chemical_list: C568108:CLDN1 protein, human; C105199:CLN3 protein, human; D062445:Claudin-1; D008562:Membrane Glycoproteins; D008565:Membrane Proteins; D018832:Molecular Chaperones; C568985:OCLN protein, human; D062793:Occludin; D014746:Vimentin; D000681:Amylases; D007610:Kallikreins