Maintenance of Differentiated Rat Hepatocytes in Primary Culture
Normal adult rat hepatocytes remained viable and functional for at least 43 days when plated on collagen-coated dishes and fed chemically defined medium supplemented with dimethyl sulfoxide (Me2SO). Hepatocytes isolated by collagenase perfusion and cultured in the presence or absence of Me2SO were (...
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| Veröffentlicht in: | Proceedings of the National Academy of Sciences - PNAS 1985-05, Vol.82 (10), p.3252-3256 |
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| container_title | Proceedings of the National Academy of Sciences - PNAS |
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| creator | Isom, Harriet C. Secott, Timothy Georgoff, Ingo Woodworth, Craig Mummaw, Jon |
| description | Normal adult rat hepatocytes remained viable and functional for at least 43 days when plated on collagen-coated dishes and fed chemically defined medium supplemented with dimethyl sulfoxide (Me2SO). Hepatocytes isolated by collagenase perfusion and cultured in the presence or absence of Me2SO were (i) examined by light and electron microscopy for morphological changes; (ii) analyzed for the production of albumin and other plasma proteins; and (iii) tested by autoradiography for DNA synthesis. Me2SO-treated cells continued to produce specific plasma proteins during the entire culture period; albumin production was consistently high (11-19 μ g/ml of culture medium per 24 hr) from day 2 to at least day 43 after plating. Ultrastructural analyses demonstrated that Me2SO-treated hepatocytes resembled those from intact liver in organization of cytoplasmic organelles and cellular junctions. The optimal concentration for observing the morphological and biochemical effects of Me2SO was 2% (vol/vol). We conclude that supplementation of chemically defined medium with Me2SO enables maintenance of differentiated hepatocytes in culture for extended periods of time. |
| doi_str_mv | 10.1073/pnas.82.10.3252 |
| format | Article |
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Hepatocytes isolated by collagenase perfusion and cultured in the presence or absence of Me2SO were (i) examined by light and electron microscopy for morphological changes; (ii) analyzed for the production of albumin and other plasma proteins; and (iii) tested by autoradiography for DNA synthesis. Me2SO-treated cells continued to produce specific plasma proteins during the entire culture period; albumin production was consistently high (11-19 μ g/ml of culture medium per 24 hr) from day 2 to at least day 43 after plating. Ultrastructural analyses demonstrated that Me2SO-treated hepatocytes resembled those from intact liver in organization of cytoplasmic organelles and cellular junctions. The optimal concentration for observing the morphological and biochemical effects of Me2SO was 2% (vol/vol). We conclude that supplementation of chemically defined medium with Me2SO enables maintenance of differentiated hepatocytes in culture for extended periods of time.</description><identifier>ISSN: 0027-8424</identifier><identifier>EISSN: 1091-6490</identifier><identifier>DOI: 10.1073/pnas.82.10.3252</identifier><identifier>PMID: 3858822</identifier><identifier>CODEN: PNASA6</identifier><language>eng</language><publisher>Washington, DC: National Academy of Sciences of the United States of America</publisher><subject>Albumins ; Albumins - metabolism ; Animal cells ; Animals ; Biological and medical sciences ; Biotechnology ; Blood proteins ; Cell culture techniques ; Cell cultures. Hybridization. Fusion ; Cell Differentiation - drug effects ; Cell lines ; Cells ; Cells, Cultured ; Collagen ; Culture Media ; Cultured cells ; Dimethyl Sulfoxide - pharmacology ; DNA - biosynthesis ; Epithelial cells ; Establishment of new cell lines, improvement of cultural methods, mass cultures ; Eukaryotic cell cultures ; Fundamental and applied biological sciences. Psychology ; Hepatocytes ; Liver - cytology ; Liver - metabolism ; Liver cells ; Methods. Procedures. Technologies ; Molecular and cellular biology ; Molecular Weight ; Proteins - metabolism ; Rats ; Tetradecanoylphorbol Acetate - pharmacology ; Time Factors</subject><ispartof>Proceedings of the National Academy of Sciences - PNAS, 1985-05, Vol.82 (10), p.3252-3256</ispartof><rights>1985 INIST-CNRS</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c556t-66a403b31c7991a655ad526b530a164fa6c3b6ea22329066aa8238dcc3cbaed33</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Uhttp://www.pnas.org/content/82/10.cover.gif</thumbnail><linktopdf>$$Uhttps://www.jstor.org/stable/pdf/25561$$EPDF$$P50$$Gjstor$$H</linktopdf><linktohtml>$$Uhttps://www.jstor.org/stable/25561$$EHTML$$P50$$Gjstor$$H</linktohtml><link.rule.ids>230,314,725,778,782,801,883,27911,27912,53778,53780,58004,58237</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=9212274$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/3858822$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Isom, Harriet C.</creatorcontrib><creatorcontrib>Secott, Timothy</creatorcontrib><creatorcontrib>Georgoff, Ingo</creatorcontrib><creatorcontrib>Woodworth, Craig</creatorcontrib><creatorcontrib>Mummaw, Jon</creatorcontrib><title>Maintenance of Differentiated Rat Hepatocytes in Primary Culture</title><title>Proceedings of the National Academy of Sciences - PNAS</title><addtitle>Proc Natl Acad Sci U S A</addtitle><description>Normal adult rat hepatocytes remained viable and functional for at least 43 days when plated on collagen-coated dishes and fed chemically defined medium supplemented with dimethyl sulfoxide (Me2SO). Hepatocytes isolated by collagenase perfusion and cultured in the presence or absence of Me2SO were (i) examined by light and electron microscopy for morphological changes; (ii) analyzed for the production of albumin and other plasma proteins; and (iii) tested by autoradiography for DNA synthesis. Me2SO-treated cells continued to produce specific plasma proteins during the entire culture period; albumin production was consistently high (11-19 μ g/ml of culture medium per 24 hr) from day 2 to at least day 43 after plating. Ultrastructural analyses demonstrated that Me2SO-treated hepatocytes resembled those from intact liver in organization of cytoplasmic organelles and cellular junctions. The optimal concentration for observing the morphological and biochemical effects of Me2SO was 2% (vol/vol). We conclude that supplementation of chemically defined medium with Me2SO enables maintenance of differentiated hepatocytes in culture for extended periods of time.</description><subject>Albumins</subject><subject>Albumins - metabolism</subject><subject>Animal cells</subject><subject>Animals</subject><subject>Biological and medical sciences</subject><subject>Biotechnology</subject><subject>Blood proteins</subject><subject>Cell culture techniques</subject><subject>Cell cultures. Hybridization. Fusion</subject><subject>Cell Differentiation - drug effects</subject><subject>Cell lines</subject><subject>Cells</subject><subject>Cells, Cultured</subject><subject>Collagen</subject><subject>Culture Media</subject><subject>Cultured cells</subject><subject>Dimethyl Sulfoxide - pharmacology</subject><subject>DNA - biosynthesis</subject><subject>Epithelial cells</subject><subject>Establishment of new cell lines, improvement of cultural methods, mass cultures</subject><subject>Eukaryotic cell cultures</subject><subject>Fundamental and applied biological sciences. Psychology</subject><subject>Hepatocytes</subject><subject>Liver - cytology</subject><subject>Liver - metabolism</subject><subject>Liver cells</subject><subject>Methods. Procedures. Technologies</subject><subject>Molecular and cellular biology</subject><subject>Molecular Weight</subject><subject>Proteins - metabolism</subject><subject>Rats</subject><subject>Tetradecanoylphorbol Acetate - pharmacology</subject><subject>Time Factors</subject><issn>0027-8424</issn><issn>1091-6490</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1985</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kMFr2zAUh0XpSNNs50GhxYdCT06kJ0uWD4WWtF0HGRtjO4tnWV4dHDtISmn--8okzbZLT-Lx-35Pj4-Qz4xOGc35bN2hnyqIw5SDgCMyZrRgqcwKekzGlEKeqgyyE3Lq_ZJSWghFR2TElVAKYExuvmHTBdthZ2zS18ldU9fW2S40GGyV_MSQPNo1ht5sg_VJ0yU_XLNCt03mmzZsnP1IPtTYevtp_07I74f7X_PHdPH9y9f57SI1QsiQSokZ5SVnJi8KhlIIrATIUnCKTGY1SsNLaRGAQ0EjjQq4qozhpkRbcT4h17u96025spWJJzps9Xp3je6x0f8nXfOk__TPmhd5Lob-bNc3rvfe2fpQZVQPKvWgUisY5kFlbJz_--OB37uL-eU-R2-wrV102PgDVgADyLOIXe2xYf9b-vcfXW_aNtiXEMmLd8kInO2ApQ-9OxAQDTP-Ch3wnk0</recordid><startdate>19850501</startdate><enddate>19850501</enddate><creator>Isom, Harriet C.</creator><creator>Secott, Timothy</creator><creator>Georgoff, Ingo</creator><creator>Woodworth, Craig</creator><creator>Mummaw, Jon</creator><general>National Academy of Sciences of the United States of America</general><general>National Acad Sciences</general><scope>IQODW</scope><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>5PM</scope></search><sort><creationdate>19850501</creationdate><title>Maintenance of Differentiated Rat Hepatocytes in Primary Culture</title><author>Isom, Harriet C. ; Secott, Timothy ; Georgoff, Ingo ; Woodworth, Craig ; Mummaw, Jon</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c556t-66a403b31c7991a655ad526b530a164fa6c3b6ea22329066aa8238dcc3cbaed33</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1985</creationdate><topic>Albumins</topic><topic>Albumins - metabolism</topic><topic>Animal cells</topic><topic>Animals</topic><topic>Biological and medical sciences</topic><topic>Biotechnology</topic><topic>Blood proteins</topic><topic>Cell culture techniques</topic><topic>Cell cultures. Hybridization. Fusion</topic><topic>Cell Differentiation - drug effects</topic><topic>Cell lines</topic><topic>Cells</topic><topic>Cells, Cultured</topic><topic>Collagen</topic><topic>Culture Media</topic><topic>Cultured cells</topic><topic>Dimethyl Sulfoxide - pharmacology</topic><topic>DNA - biosynthesis</topic><topic>Epithelial cells</topic><topic>Establishment of new cell lines, improvement of cultural methods, mass cultures</topic><topic>Eukaryotic cell cultures</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>Hepatocytes</topic><topic>Liver - cytology</topic><topic>Liver - metabolism</topic><topic>Liver cells</topic><topic>Methods. Procedures. Technologies</topic><topic>Molecular and cellular biology</topic><topic>Molecular Weight</topic><topic>Proteins - metabolism</topic><topic>Rats</topic><topic>Tetradecanoylphorbol Acetate - pharmacology</topic><topic>Time Factors</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Isom, Harriet C.</creatorcontrib><creatorcontrib>Secott, Timothy</creatorcontrib><creatorcontrib>Georgoff, Ingo</creatorcontrib><creatorcontrib>Woodworth, Craig</creatorcontrib><creatorcontrib>Mummaw, Jon</creatorcontrib><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</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>Isom, Harriet C.</au><au>Secott, Timothy</au><au>Georgoff, Ingo</au><au>Woodworth, Craig</au><au>Mummaw, Jon</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Maintenance of Differentiated Rat Hepatocytes in Primary Culture</atitle><jtitle>Proceedings of the National Academy of Sciences - PNAS</jtitle><addtitle>Proc Natl Acad Sci U S A</addtitle><date>1985-05-01</date><risdate>1985</risdate><volume>82</volume><issue>10</issue><spage>3252</spage><epage>3256</epage><pages>3252-3256</pages><issn>0027-8424</issn><eissn>1091-6490</eissn><coden>PNASA6</coden><abstract>Normal adult rat hepatocytes remained viable and functional for at least 43 days when plated on collagen-coated dishes and fed chemically defined medium supplemented with dimethyl sulfoxide (Me2SO). Hepatocytes isolated by collagenase perfusion and cultured in the presence or absence of Me2SO were (i) examined by light and electron microscopy for morphological changes; (ii) analyzed for the production of albumin and other plasma proteins; and (iii) tested by autoradiography for DNA synthesis. Me2SO-treated cells continued to produce specific plasma proteins during the entire culture period; albumin production was consistently high (11-19 μ g/ml of culture medium per 24 hr) from day 2 to at least day 43 after plating. Ultrastructural analyses demonstrated that Me2SO-treated hepatocytes resembled those from intact liver in organization of cytoplasmic organelles and cellular junctions. The optimal concentration for observing the morphological and biochemical effects of Me2SO was 2% (vol/vol). We conclude that supplementation of chemically defined medium with Me2SO enables maintenance of differentiated hepatocytes in culture for extended periods of time.</abstract><cop>Washington, DC</cop><pub>National Academy of Sciences of the United States of America</pub><pmid>3858822</pmid><doi>10.1073/pnas.82.10.3252</doi><tpages>5</tpages><oa>free_for_read</oa></addata></record> |
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| ispartof | Proceedings of the National Academy of Sciences - PNAS, 1985-05, Vol.82 (10), p.3252-3256 |
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| subjects | Albumins Albumins - metabolism Animal cells Animals Biological and medical sciences Biotechnology Blood proteins Cell culture techniques Cell cultures. Hybridization. Fusion Cell Differentiation - drug effects Cell lines Cells Cells, Cultured Collagen Culture Media Cultured cells Dimethyl Sulfoxide - pharmacology DNA - biosynthesis Epithelial cells Establishment of new cell lines, improvement of cultural methods, mass cultures Eukaryotic cell cultures Fundamental and applied biological sciences. Psychology Hepatocytes Liver - cytology Liver - metabolism Liver cells Methods. Procedures. Technologies Molecular and cellular biology Molecular Weight Proteins - metabolism Rats Tetradecanoylphorbol Acetate - pharmacology Time Factors |
| title | Maintenance of Differentiated Rat Hepatocytes in Primary Culture |
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