Isolation, maintenance, and characterization of human pancreatic islet tumor cells expressing vasoactive intestinal peptide.
Tissue from a vasoactive intestinal peptide (VIP)-secreting human tumor has been used to establish and characterize human neuroendocrine primary cell cultures from which permanent, clone-derived cell lines have been established. Viable cells were obtained by enzymatic and mechanical dissociation of freshly resected pancreatic islet tumor and hepatic metastatic tumor tissues. Aliquots of tumor cells were established ex vivo under culture conditions including porous substrata coated with type IV collagen and laminin and a low serum, hormonally defined culture medium. The small (<10 microm) rounded, grape-like cells had a very slow growth rate of doubling times estimated at several weeks or more. After several passages, morphologically uniform cells were derived that strongly expressed neuroendocrine markers of synaptophysin and synaptobrevin. Although chromogranin A and VIP had somewhat weaker expression, both demonstrated phorbol ester-stimulated secretion. The morphologic and secretory properties were maintained by the cells for nearly 2 years in culture. The establishment of this novel VIP-secreting human neuroendocrine cell line (HuNET) makes available a culture model with which to study a transformed version of this pancreatic islet cell type and offers approaches by which to establish islet tumor cell lines.
Tillotson
L G
LG
Department of Medicine, University of North Carolina School of Medicine, Chapel Hill 27599-7038, USA. lgtgi@med.unc.edu
Lodestro
C
C
Höcker
M
M
Wiedenmann
B
B
Newcomer
C E
CE
Reid
L M
LM
D000328:Adult; D018273:Carcinoma, Islet Cell / Q000378:metabolism* / Q000556:secondary; D002469:Cell Separation; D053379:Chromogranin A; D002864:Chromogranins / Q000378:metabolism; D015925:Cryopreservation; D019084:Fluorescent Antibody Technique, Indirect; D006801:Humans; D007124:Immunoenzyme Techniques; D008113:Liver Neoplasms / Q000378:metabolism* / Q000556:secondary; D008297:Male; D010190:Pancreatic Neoplasms / Q000378:metabolism* / Q000473:pathology; D016708:Synaptophysin / Q000378:metabolism; D014407:Tumor Cells, Cultured; D014660:Vasoactive Intestinal Peptide / Q000378:metabolism*
D053379:Chromogranin A; D002864:Chromogranins; D016708:Synaptophysin; D014660:Vasoactive Intestinal Peptide