Abstract:
:Cultivation of human parotid glands in serum-free medium (Ca(2+) concentration, 0.2 mM) with growth supplements resulted in isolation of a homogeneous population of epithelial cells without any mesenchymal cells. The isolated cells showed an undifferentiated phenotype with scant cytoplasmic organelles, and low levels of alpha-amylase expression. The cells remained viable and undifferentiated for up to 24 passages when subcultured at 80% confluence in 0.2 mM Ca(2+) medium with a 1:3 split ratios. There was little cell-cell contact. A Ca(2+) switch from 0.2 to 1 mM induced cell-cell contact with translocation of desmosomal proteins from the cytoplasm to the cell membrane, and sequential differentiation of serous acinar cells with a glandular arrangement, well-developed cytoplasmic organelles and an increased level of alpha-amylase expression. These morphological changes and desmosome assembly were blocked by treatment with non-specific PKC inhibitor. Moreover, the addition of PKC activator, tetradecanoylphorbol 13-acetate (TPA), to 0.2 mM Ca(2+) medium caused transient assembly of desmosome-like structure, but did not induce cell-cell contact or morphological differentiation. Cultivation of the cells in 1.5 mM Ca(2+) medium resulted in increased stratification of the cells and reduced alpha-amylase expression. These findings provide the first demonstration that continuous cultivation in 1.0 mM Ca(2+) medium is required for cellular differentiation of salivary gland acinar cells, and maintenance of the differentiated state.
journal_name
J Cell Physioljournal_title
Journal of cellular physiologyauthors
Hiraki A,Shirasuna K,Ikari T,Shinohara M,Garrod DRdoi
10.1002/jcp.10149keywords:
subject
Has Abstractpub_date
2002-10-01 00:00:00pages
55-63issue
1eissn
0021-9541issn
1097-4652journal_volume
193pub_type
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