Insulin Synthesis in a Clonal Cell Line of Simian Virus 40-Transformed Hamster Pancreatic Beta Cells

A clonal hamster beta cell line (HIT) was established by simian virus 40 transformation of Syrian hamster pancreatic islet cells. Cytoplasmic insulin was detected in all cells by indirect fluorescent antibody staining, and membrane-bound secretory granules were observed ultrastructurally. Acidified-...

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Bibliographic Details
Published in:Proceedings of the National Academy of Sciences - PNAS 1981-07, Vol.78 (7), p.4339-4343
Main Authors: Santerre, Robert F., Cook, Roland A., Rae Marie D. Crisel, Sharp, John D., Schmidt, Robert J., Williams, Daniel C., Wilson, Christine P.
Format: Article
Language:English
Subjects:
Animals
Antibodies
Beta cells
Cell culture techniques
Cell Line
Cell lines
Cell Transformation, Viral
Cells
Clone Cells
Cricetinae
Cultured cells
Endocrine cells
Gels
Glucagon - pharmacology
Glucose - pharmacology
Insulin
Insulin - biosynthesis
Insulin - metabolism
Insulin Secretion
Islet cells
Islets of Langerhans - metabolism
Mesocricetus
Secretory Rate - drug effects
Simian virus 40
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Summary:A clonal hamster beta cell line (HIT) was established by simian virus 40 transformation of Syrian hamster pancreatic islet cells. Cytoplasmic insulin was detected in all cells by indirect fluorescent antibody staining, and membrane-bound secretory granules were observed ultrastructurally. Acidified-ethanol extracts of HIT cell cultures contained hamster insulin as determined by radioimmunoassay, radioreceptor assay, and bioassay. One subclone at passage 39 contained 2.6 μ g of insulin per mg of cell protein. [3H]Leucine-labeled HIT insulin and proinsulin were identical to islet-derived proteins when compared by NaDodSO4/polyacrylamide gel electrophoresis of immunoprecipitates. HIT cell insulin secretion was stimulated by glucose, glucagon, and 3-isobutyl-1-methylxanthine. Insulin secretion at optimal glucose concentration (7.5 mM) was 2.4 milliunits per 106cells per hr. Somatostatin and dexamethasone markedly inhibited HIT insulin secretion. The HIT cell line represents a unique in vitro system for studying beta cell metabolism and insulin biosynthesis.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.78.7.4339