TY - JOUR
T1 - Transfection with SV40 gene of human pancreatic endocrine cells
AU - Soldevila, Gloria
AU - Buscema, Massimo
AU - Marini, Vittoria
AU - Sutton, Robert
AU - James, Roger F.L.
AU - Bloom, Stephen R.
AU - Robertson, R. Paul
AU - Mirakian, Rita
AU - Pujol-Borrell, Ricardo
AU - Bottazzo, Gian Franco
PY - 1991/1/1
Y1 - 1991/1/1
N2 - At present, only islet cell lines of animal origin have been successfully generated (e.g. RIN, HIT). A fully differentiated human β 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-γ and modulated by TNF-α. 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. © 1991.
AB - At present, only islet cell lines of animal origin have been successfully generated (e.g. RIN, HIT). A fully differentiated human β 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-γ and modulated by TNF-α. 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. © 1991.
U2 - 10.1016/0896-8411(91)90154-5
DO - 10.1016/0896-8411(91)90154-5
M3 - Article
SN - 0896-8411
VL - 4
SP - 381
EP - 396
JO - Journal of Autoimmunity
JF - Journal of Autoimmunity
IS - 3
ER -