Development and Characterization of SV40 Immortalized Rat Submandibular Acinar Cell Lines

Rat submandibular salivary gland acinar cells were transfected by CaPO4precipitation using a plasmid containing a replication-defective simian virus (SV40) genome. Out of 27 clonal cell lines, two were shown to have moderate to high levels of cytodifferentiation and salivary gland acinar cell functi...

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Bibliographic Details
Published in:In vitro cellular & developmental biology. Animal 1997-03, Vol.33 (3), p.164-173
Main Authors: D. O. Quissell, K. A. Barzen, Gruenert, D. C., R. S. Redman, J. M. Camden, Turner, J. T.
Format: Article
Language:English
Subjects:
Acinar cells
Agonists
Animals
Calcium - metabolism
Cell growth
Cell Line, Transformed
Cell lines
Cellular Models
Cultured cells
Cyclic AMP - biosynthesis
Epithelial cells
Fluorescent Antibody Technique, Indirect
Inositol Phosphates - metabolism
Male
Microscopy, Electron
Phenylephrine
Plasmids
Rats
Rats, Sprague-Dawley
Receptors
Salivary glands
Simian virus 40 - genetics
Submandibular Gland - cytology
Submandibular Gland - drug effects
Transfection
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Summary:Rat submandibular salivary gland acinar cells were transfected by CaPO4precipitation using a plasmid containing a replication-defective simian virus (SV40) genome. Out of 27 clonal cell lines, two were shown to have moderate to high levels of cytodifferentiation and salivary gland acinar cell function. Functional studies with the two cell lines indicated that the$\beta-adrenergic$agonist, isoproterenol, vasoactive intestinal peptide, and prostaglandin E1were effective activators of intracellular cyclic AMP production. Epinephrine, norepinephrine, phenylephrine, acetylcholine, and$P_{2U}-purinoceptor$agonists were effective in increasing inositol phosphate production and intracellular free calcium levels, whereas substance P was without effect. Utilizing indirect immunofluorescence analysis, both cell lines were shown to express glutamine/ glutamic acid-rich proteins, a submandibular acinar cell specific secretory protein family. Electron microscopic evaluation documented the maintenance of tripartite junctional complexes, cellular polarization, and the presence of moderate amounts of secretory granules and rough endoplasmic reticulum. The two cell lines had doubling times of 25 h.
ISSN:1071-2690
1543-706X
DOI:10.1007/s11626-997-0137-8