Transfection with SV40 gene of human pancreatic endocrine cells.
At present, only islet cell lines of animal origin have been successfully generated (e.g. RIN, HIT). A fully differentiated human beta cell line would be advantageous for diabetes research. We now report the generation of a human endocrine pancreatic cell line obtained by transfection using a plasmid containing the early region of SV40 viral DNA. Viral integration and transcription was assessed by Southern and Northern blotting. This cell line has been growing continuously for more than 2 years and maintains several of the characteristics of the parental cells from which they were generated. The presence of Neuron Specific Enolase, Protein Gene Product 9.5, cytokeratin, microvilli, cytoplasmic electrodense granules and the secretion of insulin, glucagon and somatostatin supports the neuroendocrine origin of this cell line. However, hormone production progressively decreased and finally stopped at passage 8. Flow cytometric analysis showed that HLA expression in this cell line is readily induced by IFN-gamma and modulated by TNF-alpha. The establishment of this human endocrine cell line indicates the feasibility of immortalizing human islets by transfection with viral oncogenes. To obtain a fully differentiated cell line it may be necessary to use other DNA constructs which immortalize the cells without fully transforming their phenotype.
Department of Immunology, University College and Middlesex School of Medicine, London, UK.
G
G
Soldevila
M
M
Buscema
V
V
Marini
R
R
Sutton
R F
RF
James
S R
SR
Bloom
R P
RP
Robertson
R
R
Mirakian
R
R
Pujol-Borrell
G F
GF
Bottazzo
D000957:Antigens, Viral, Tumor / Q000235:genetics; D002461:Cell Line, Transformed; D004279:DNA, Viral / Q000235:genetics; D005934:Glucagon / Q000378:metabolism; D006680:HLA Antigens; D006801:Humans; D007150:Immunohistochemistry; D007328:Insulin / Q000378:metabolism; D007515:Islets of Langerhans / Q000166:cytology* / Q000276:immunology / Q000378:metabolism; D013539:Simian virus 40 / Q000235:genetics* / Q000276:immunology; D013004:Somatostatin / Q000378:metabolism; D014162:Transfection
D000957:Antigens, Viral, Tumor; D004279:DNA, Viral; D006680:HLA Antigens; D007328:Insulin; D013004:Somatostatin; D005934:Glucagon