{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2023,10,17]],"date-time":"2023-10-17T01:55:24Z","timestamp":1697507724611},"reference-count":0,"publisher":"American Diabetes Association","issue":"10","content-domain":{"domain":["diabetesjournals.org"],"crossmark-restriction":true},"short-container-title":[],"published-print":{"date-parts":[[1999,10,1]]},"abstract":"<jats:p>Type 2 diabetes is characterized by islet amyloid deposits, which are primarily composed of the amyloidogenic human form of islet amyloid polypeptide (IAPP, amylin). The mechanism of islet amyloido-genesis is not known, but other products (e.g., apolipoprotein E and perlecan) contained within islet amyloid may be necessary. Because rodent IAPP does not form islet amyloid, the currently available beta-cell lines are not useful for studying processes involved in amyloid formation. To develop a suitable in vitro cell system for the study of islet amyloid formation, we generated two new beta-cell lines that express the amyloidogenic human IAPP. We did this by crossbreeding human IAPP transgenic mice with RIP-Tag mice that develop islet tumors and then culturing one of these islet tumors from two separate offspring of this cross. The resultant 2350-2C0 and 2511 cell lines produce human as well as mouse IAPP-like immunoreactivity (IAPP-LI) and immunoreactive insulin (IRI). Incubation of both these cell lines with 16.7 mmol\/l glucose resulted in a two- to fourfold increase in human IAPP-LI, mouse IAPP-LI, and IRI secretion compared with 1.67 mmol\/l glucose and the combination of 16.7 mmol\/l glucose and 10 mmol\/l arginine, 0.1 mmol\/l 3-isobutyl-1-methylxanthine (IBMX), and 5 micromol\/l carbachol induced a &amp;gt;50-fold increase in the release of these peptides. The omission of calcium from the above secretagogue cocktail reduced secretion of all three peptides to only two- to sixfold higher than the 16.7 mmol\/l glucose condition. Perifusion with 16.7 mmol\/l glucose plus 0.1 mmol\/l IBMX caused a biphasic secretion of human IAPP-LI and mouse IAPP-LI, as well as IRI, in both cell lines, with the peak of the first phase being five- to sixfold higher than the prestimulated 1.67 mmol\/l glucose condition. Immunoelectron microscopic inspection of both 2350-2C0 and 2511 cells after 7 days of culture did not reveal the presence of amyloid fibrils, suggesting the need for other critical components. We conclude that we have established two novel beta-cell lines that produce and secrete human IAPP in a regulated manner. These cell lines will be a useful tool to investigate the secretion of human IAPP as well as the necessity of other components for islet amyloid formation.<\/jats:p>","DOI":"10.2337\/diabetes.48.10.1962","type":"journal-article","created":{"date-parts":[[2007,3,6]],"date-time":"2007-03-06T19:04:22Z","timestamp":1173207862000},"page":"1962-1970","update-policy":"http:\/\/dx.doi.org\/10.2337\/ada-journal-policies","source":"Crossref","is-referenced-by-count":13,"title":["Two novel immortal pancreatic beta-cell lines expressing and secreting human islet amyloid polypeptide do not spontaneously develop islet amyloid."],"prefix":"10.2337","volume":"48","author":[{"given":"S","family":"Andrikopoulos","sequence":"first","affiliation":[{"name":"Department of Medicine, University of Washington, Seattle, USA."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"C B","family":"Verchere","sequence":"additional","affiliation":[{"name":"Department of Medicine, University of Washington, Seattle, USA."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"J C","family":"Teague","sequence":"additional","affiliation":[{"name":"Department of Medicine, University of Washington, Seattle, USA."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"W M","family":"Howell","sequence":"additional","affiliation":[{"name":"Department of Medicine, University of Washington, Seattle, USA."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"W Y","family":"Fujimoto","sequence":"additional","affiliation":[{"name":"Department of Medicine, University of Washington, Seattle, USA."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"T N","family":"Wight","sequence":"additional","affiliation":[{"name":"Department of Medicine, University of Washington, Seattle, USA."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"S E","family":"Kahn","sequence":"additional","affiliation":[{"name":"Department of Medicine, University of Washington, Seattle, USA."}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1167","container-title":["Diabetes"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/journals.org\/diabetes\/diabetes\/article-pdf\/48\/10\/1962\/363813\/10512360.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/diabetesjournals.org\/diabetes\/article-pdf\/48\/10\/1962\/363813\/10512360.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,11,2]],"date-time":"2022-11-02T16:46:09Z","timestamp":1667407569000},"score":1,"resource":{"primary":{"URL":"https:\/\/diabetesjournals.org\/diabetes\/article\/48\/10\/1962\/11422\/Two-novel-immortal-pancreatic-beta-cell-lines"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[1999,10,1]]},"references-count":0,"journal-issue":{"issue":"10","published-print":{"date-parts":[[1999,10,1]]}},"URL":"https:\/\/doi.org\/10.2337\/diabetes.48.10.1962","relation":{},"ISSN":["0012-1797","1939-327X"],"issn-type":[{"value":"0012-1797","type":"print"},{"value":"1939-327X","type":"electronic"}],"subject":[],"published":{"date-parts":[[1999,10,1]]}}}