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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| Veröffentlicht in: | Proceedings of the National Academy of Sciences - PNAS 1981-07, Vol.78 (7), p.4339-4343 |
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| container_title | Proceedings of the National Academy of Sciences - PNAS |
| container_volume | 78 |
| creator | Santerre, Robert F. Cook, Roland A. Rae Marie D. Crisel Sharp, John D. Schmidt, Robert J. Williams, Daniel C. Wilson, Christine P. |
| description | 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. |
| doi_str_mv | 10.1073/pnas.78.7.4339 |
| format | Article |
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Crisel ; Sharp, John D. ; Schmidt, Robert J. ; Williams, Daniel C. ; Wilson, Christine P.</creator><creatorcontrib>Santerre, Robert F. ; Cook, Roland A. ; Rae Marie D. Crisel ; Sharp, John D. ; Schmidt, Robert J. ; Williams, Daniel C. ; Wilson, Christine P.</creatorcontrib><description>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.</description><identifier>ISSN: 0027-8424</identifier><identifier>EISSN: 1091-6490</identifier><identifier>DOI: 10.1073/pnas.78.7.4339</identifier><identifier>PMID: 6270673</identifier><language>eng</language><publisher>United States: National Academy of Sciences of the United States of America</publisher><subject>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</subject><ispartof>Proceedings of the National Academy of Sciences - PNAS, 1981-07, Vol.78 (7), p.4339-4343</ispartof><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c553t-ac3ee10d70b8d69f2348d793300f606903240fe376b5e19e1f629bd5aba2bb253</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Uhttp://www.pnas.org/content/78/7.cover.gif</thumbnail><linktopdf>$$Uhttps://www.jstor.org/stable/pdf/10608$$EPDF$$P50$$Gjstor$$H</linktopdf><linktohtml>$$Uhttps://www.jstor.org/stable/10608$$EHTML$$P50$$Gjstor$$H</linktohtml><link.rule.ids>230,314,727,780,784,803,885,27924,27925,53791,53793,58017,58250</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/6270673$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Santerre, Robert F.</creatorcontrib><creatorcontrib>Cook, Roland A.</creatorcontrib><creatorcontrib>Rae Marie D. Crisel</creatorcontrib><creatorcontrib>Sharp, John D.</creatorcontrib><creatorcontrib>Schmidt, Robert J.</creatorcontrib><creatorcontrib>Williams, Daniel C.</creatorcontrib><creatorcontrib>Wilson, Christine P.</creatorcontrib><title>Insulin Synthesis in a Clonal Cell Line of Simian Virus 40-Transformed Hamster Pancreatic Beta Cells</title><title>Proceedings of the National Academy of Sciences - PNAS</title><addtitle>Proc Natl Acad Sci U S A</addtitle><description>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.</description><subject>Animals</subject><subject>Antibodies</subject><subject>Beta cells</subject><subject>Cell culture techniques</subject><subject>Cell Line</subject><subject>Cell lines</subject><subject>Cell Transformation, Viral</subject><subject>Cells</subject><subject>Clone Cells</subject><subject>Cricetinae</subject><subject>Cultured cells</subject><subject>Endocrine cells</subject><subject>Gels</subject><subject>Glucagon - pharmacology</subject><subject>Glucose - pharmacology</subject><subject>Insulin</subject><subject>Insulin - biosynthesis</subject><subject>Insulin - metabolism</subject><subject>Insulin Secretion</subject><subject>Islet cells</subject><subject>Islets of Langerhans - metabolism</subject><subject>Mesocricetus</subject><subject>Secretory Rate - drug effects</subject><subject>Simian virus 40</subject><issn>0027-8424</issn><issn>1091-6490</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1981</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFUc9rFDEYDaLUbfXqQRBy6m3GL8lMMjl40MXawkILrV5DZiZjUzLJmmSk_e-dcddlBcFT-Hg_8h4PoTcESgKCvd96nUrRlKKsGJPP0IqAJAWvJDxHKwAqiqai1Ut0mtIDAMi6gRN0wqkALtgK9Vc-Tc56fPvk871JNuH50HjtgtcOr41zeGO9wWHAt3a02uNvNk4JV1DcRe3TEOJoenypx5RNxDfad9HobDv8yWT92yC9Qi8G7ZJ5vX_P0NeLz3fry2Jz_eVq_XFTdHXNcqE7ZgyBXkDb9FwOlFVNLyRjAAMHLoHRCgbDBG9rQ6QhA6ey7Wvdatq2tGZn6MPOdzu1c6jO-By1U9toRx2fVNBW_Y14e6--h5-KESmaRX--18fwYzIpq9Gmbm6gvQlTUoJxIWpK_kskdUXl3GkmljtiF0NK0QyHMATUsp9a9lOiUUIt-82Cd8cVDvT9YEc_L7o_6EGvhsm5bB7zkdE_iTP-doc_pBziUSwODfsFab-3kQ</recordid><startdate>19810701</startdate><enddate>19810701</enddate><creator>Santerre, Robert F.</creator><creator>Cook, Roland A.</creator><creator>Rae Marie D. Crisel</creator><creator>Sharp, John D.</creator><creator>Schmidt, Robert J.</creator><creator>Williams, Daniel C.</creator><creator>Wilson, Christine P.</creator><general>National Academy of Sciences of the United States of America</general><general>National Acad Sciences</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7U9</scope><scope>H94</scope><scope>7X8</scope><scope>5PM</scope></search><sort><creationdate>19810701</creationdate><title>Insulin Synthesis in a Clonal Cell Line of Simian Virus 40-Transformed Hamster Pancreatic Beta Cells</title><author>Santerre, Robert F. ; Cook, Roland A. ; Rae Marie D. Crisel ; Sharp, John D. ; Schmidt, Robert J. ; Williams, Daniel C. ; Wilson, Christine P.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c553t-ac3ee10d70b8d69f2348d793300f606903240fe376b5e19e1f629bd5aba2bb253</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1981</creationdate><topic>Animals</topic><topic>Antibodies</topic><topic>Beta cells</topic><topic>Cell culture techniques</topic><topic>Cell Line</topic><topic>Cell lines</topic><topic>Cell Transformation, Viral</topic><topic>Cells</topic><topic>Clone Cells</topic><topic>Cricetinae</topic><topic>Cultured cells</topic><topic>Endocrine cells</topic><topic>Gels</topic><topic>Glucagon - pharmacology</topic><topic>Glucose - pharmacology</topic><topic>Insulin</topic><topic>Insulin - biosynthesis</topic><topic>Insulin - metabolism</topic><topic>Insulin Secretion</topic><topic>Islet cells</topic><topic>Islets of Langerhans - metabolism</topic><topic>Mesocricetus</topic><topic>Secretory Rate - drug effects</topic><topic>Simian virus 40</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Santerre, Robert F.</creatorcontrib><creatorcontrib>Cook, Roland A.</creatorcontrib><creatorcontrib>Rae Marie D. Crisel</creatorcontrib><creatorcontrib>Sharp, John D.</creatorcontrib><creatorcontrib>Schmidt, Robert J.</creatorcontrib><creatorcontrib>Williams, Daniel C.</creatorcontrib><creatorcontrib>Wilson, Christine P.</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Virology and AIDS Abstracts</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Proceedings of the National Academy of Sciences - PNAS</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Santerre, Robert F.</au><au>Cook, Roland A.</au><au>Rae Marie D. Crisel</au><au>Sharp, John D.</au><au>Schmidt, Robert J.</au><au>Williams, Daniel C.</au><au>Wilson, Christine P.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Insulin Synthesis in a Clonal Cell Line of Simian Virus 40-Transformed Hamster Pancreatic Beta Cells</atitle><jtitle>Proceedings of the National Academy of Sciences - PNAS</jtitle><addtitle>Proc Natl Acad Sci U S A</addtitle><date>1981-07-01</date><risdate>1981</risdate><volume>78</volume><issue>7</issue><spage>4339</spage><epage>4343</epage><pages>4339-4343</pages><issn>0027-8424</issn><eissn>1091-6490</eissn><abstract>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.</abstract><cop>United States</cop><pub>National Academy of Sciences of the United States of America</pub><pmid>6270673</pmid><doi>10.1073/pnas.78.7.4339</doi><tpages>5</tpages><oa>free_for_read</oa></addata></record> |
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| identifier | ISSN: 0027-8424 |
| ispartof | Proceedings of the National Academy of Sciences - PNAS, 1981-07, Vol.78 (7), p.4339-4343 |
| issn | 0027-8424 1091-6490 |
| language | eng |
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| source | MEDLINE; JSTOR Archive Collection A-Z Listing; PubMed Central; Alma/SFX Local Collection; Free Full-Text Journals in Chemistry |
| 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 |
| title | Insulin Synthesis in a Clonal Cell Line of Simian Virus 40-Transformed Hamster Pancreatic Beta Cells |
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