Establishment and characterization of conditionally immortalized endothelial cell lines from the thoracic duct and inferior vena cava of tsA58/EGFP double-transgenic rats
The basic biology of blood vascular endothelial cells has been well documented. However, little is known about that of lymphatic endothelial cells, despite their importance under normal and pathological conditions. The lack of a lymphatic endothelial cell line has hampered progress in this field. Th...
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| Veröffentlicht in: | Cell and tissue research 2006-12, Vol.326 (3), p.749-758 |
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| creator | Matsuo, Mitsuhiro Koizumi, Keiichi Yamada, Sanae Tomi, Masatoshi Takahashi, Ri-ichi Ueda, Masatsugu Terasaki, Tetsuya Obinata, Masuo Hosoya, Ken-ichi Ohtani, Osamu Saiki, Ikuo |
| description | The basic biology of blood vascular endothelial cells has been well documented. However, little is known about that of lymphatic endothelial cells, despite their importance under normal and pathological conditions. The lack of a lymphatic endothelial cell line has hampered progress in this field. The objective of this study has been to establish and characterize lymphatic and venous endothelial cell lines derived from newly developed tsA58/EGFP transgenic rats harboring the temperature-sensitive simian virus 40 (SV40) large T-antigen and enhanced green fluorescent protein (EGFP). Endothelial cells were isolated from the transgenic rats by intraluminal enzymatic digestion. The cloned cell lines were named TR-LE (temperature-sensitive rat lymphatic endothelial cells from thoracic duct) and TR-BE (temperature-sensitive rat blood-vessel endothelial cells from inferior vena cava), respectively, and cultured on fibronectin-coated dishes in HuMedia-EG2 supplemented with 20% fetal bovine serum and Endothelial Mitogen at a permissive temperature, 33 degrees C. A temperature shift to 37 degrees C resulted in a decrease in proliferation with degradation of the large T-antigen and cleavage of poly (ADP-ribose) polymerase. TR-LE cells expressed lymphatic endothelial markers VEGFR-3 (vascular endothelial growth factor receptor), LYVE-1 (a lymphatic endothelial receptor), Prox-1 (a homeobox gene product), and podoplanin (a glomerular podocyte membrane mucoprotein), together with endothelial markers CD31, Tie-2, and VEGFR-2, whereas TR-BE cells expressed CD31, Tie-2, and VEGFR-2, but no lymphatic endothelial markers. Thus, these conditionally immortalized and EGFP-expressing lymphatic and vascular endothelial cell lines might represent an important tool for the study of endothelial cell functions in vitro. |
| doi_str_mv | 10.1007/s00441-006-0229-x |
| format | Article |
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However, little is known about that of lymphatic endothelial cells, despite their importance under normal and pathological conditions. The lack of a lymphatic endothelial cell line has hampered progress in this field. The objective of this study has been to establish and characterize lymphatic and venous endothelial cell lines derived from newly developed tsA58/EGFP transgenic rats harboring the temperature-sensitive simian virus 40 (SV40) large T-antigen and enhanced green fluorescent protein (EGFP). Endothelial cells were isolated from the transgenic rats by intraluminal enzymatic digestion. The cloned cell lines were named TR-LE (temperature-sensitive rat lymphatic endothelial cells from thoracic duct) and TR-BE (temperature-sensitive rat blood-vessel endothelial cells from inferior vena cava), respectively, and cultured on fibronectin-coated dishes in HuMedia-EG2 supplemented with 20% fetal bovine serum and Endothelial Mitogen at a permissive temperature, 33 degrees C. A temperature shift to 37 degrees C resulted in a decrease in proliferation with degradation of the large T-antigen and cleavage of poly (ADP-ribose) polymerase. TR-LE cells expressed lymphatic endothelial markers VEGFR-3 (vascular endothelial growth factor receptor), LYVE-1 (a lymphatic endothelial receptor), Prox-1 (a homeobox gene product), and podoplanin (a glomerular podocyte membrane mucoprotein), together with endothelial markers CD31, Tie-2, and VEGFR-2, whereas TR-BE cells expressed CD31, Tie-2, and VEGFR-2, but no lymphatic endothelial markers. Thus, these conditionally immortalized and EGFP-expressing lymphatic and vascular endothelial cell lines might represent an important tool for the study of endothelial cell functions in vitro.</description><identifier>ISSN: 0302-766X</identifier><identifier>EISSN: 1432-0878</identifier><identifier>DOI: 10.1007/s00441-006-0229-x</identifier><identifier>PMID: 16773315</identifier><language>eng</language><publisher>Germany: Springer Nature B.V</publisher><subject>Animals ; Animals, Genetically Modified ; Antigens, Viral, Tumor - genetics ; Biomarkers - analysis ; Cell Culture Techniques ; Cell Line, Transformed ; Cell Proliferation ; Cell Transformation, Viral ; Cells ; Clone Cells ; Endothelial Cells - cytology ; Endothelial Cells - metabolism ; Endothelial Cells - physiology ; Endothelial Cells - ultrastructure ; Endothelium, Lymphatic - cytology ; Endothelium, Vascular - cytology ; Enzymes ; Fluorescent Dyes - metabolism ; Gene Expression Profiling ; Green Fluorescent Proteins - metabolism ; Lymphatic system ; Pulmonary arteries ; Rats ; Rodents ; Simian virus 40 - physiology ; Temperature ; Thoracic Duct - cytology ; Vena Cava, Inferior - cytology</subject><ispartof>Cell and tissue research, 2006-12, Vol.326 (3), p.749-758</ispartof><rights>Springer-Verlag 2006</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c392t-caf8d76476ab1d2bcee2215e7f19df46384fdb08bd994e256f72fcbb4a2908e73</citedby><cites>FETCH-LOGICAL-c392t-caf8d76476ab1d2bcee2215e7f19df46384fdb08bd994e256f72fcbb4a2908e73</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,777,781,27905,27906</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/16773315$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Matsuo, Mitsuhiro</creatorcontrib><creatorcontrib>Koizumi, Keiichi</creatorcontrib><creatorcontrib>Yamada, Sanae</creatorcontrib><creatorcontrib>Tomi, Masatoshi</creatorcontrib><creatorcontrib>Takahashi, Ri-ichi</creatorcontrib><creatorcontrib>Ueda, Masatsugu</creatorcontrib><creatorcontrib>Terasaki, Tetsuya</creatorcontrib><creatorcontrib>Obinata, Masuo</creatorcontrib><creatorcontrib>Hosoya, Ken-ichi</creatorcontrib><creatorcontrib>Ohtani, Osamu</creatorcontrib><creatorcontrib>Saiki, Ikuo</creatorcontrib><title>Establishment and characterization of conditionally immortalized endothelial cell lines from the thoracic duct and inferior vena cava of tsA58/EGFP double-transgenic rats</title><title>Cell and tissue research</title><addtitle>Cell Tissue Res</addtitle><description>The basic biology of blood vascular endothelial cells has been well documented. However, little is known about that of lymphatic endothelial cells, despite their importance under normal and pathological conditions. The lack of a lymphatic endothelial cell line has hampered progress in this field. The objective of this study has been to establish and characterize lymphatic and venous endothelial cell lines derived from newly developed tsA58/EGFP transgenic rats harboring the temperature-sensitive simian virus 40 (SV40) large T-antigen and enhanced green fluorescent protein (EGFP). Endothelial cells were isolated from the transgenic rats by intraluminal enzymatic digestion. The cloned cell lines were named TR-LE (temperature-sensitive rat lymphatic endothelial cells from thoracic duct) and TR-BE (temperature-sensitive rat blood-vessel endothelial cells from inferior vena cava), respectively, and cultured on fibronectin-coated dishes in HuMedia-EG2 supplemented with 20% fetal bovine serum and Endothelial Mitogen at a permissive temperature, 33 degrees C. A temperature shift to 37 degrees C resulted in a decrease in proliferation with degradation of the large T-antigen and cleavage of poly (ADP-ribose) polymerase. TR-LE cells expressed lymphatic endothelial markers VEGFR-3 (vascular endothelial growth factor receptor), LYVE-1 (a lymphatic endothelial receptor), Prox-1 (a homeobox gene product), and podoplanin (a glomerular podocyte membrane mucoprotein), together with endothelial markers CD31, Tie-2, and VEGFR-2, whereas TR-BE cells expressed CD31, Tie-2, and VEGFR-2, but no lymphatic endothelial markers. Thus, these conditionally immortalized and EGFP-expressing lymphatic and vascular endothelial cell lines might represent an important tool for the study of endothelial cell functions in vitro.</description><subject>Animals</subject><subject>Animals, Genetically Modified</subject><subject>Antigens, Viral, Tumor - genetics</subject><subject>Biomarkers - analysis</subject><subject>Cell Culture Techniques</subject><subject>Cell Line, Transformed</subject><subject>Cell Proliferation</subject><subject>Cell Transformation, Viral</subject><subject>Cells</subject><subject>Clone Cells</subject><subject>Endothelial Cells - cytology</subject><subject>Endothelial Cells - metabolism</subject><subject>Endothelial Cells - physiology</subject><subject>Endothelial Cells - ultrastructure</subject><subject>Endothelium, Lymphatic - cytology</subject><subject>Endothelium, Vascular - cytology</subject><subject>Enzymes</subject><subject>Fluorescent Dyes - metabolism</subject><subject>Gene Expression Profiling</subject><subject>Green Fluorescent Proteins - metabolism</subject><subject>Lymphatic system</subject><subject>Pulmonary arteries</subject><subject>Rats</subject><subject>Rodents</subject><subject>Simian virus 40 - physiology</subject><subject>Temperature</subject><subject>Thoracic Duct - cytology</subject><subject>Vena Cava, Inferior - cytology</subject><issn>0302-766X</issn><issn>1432-0878</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2006</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>GNUQQ</sourceid><recordid>eNpdkc1KHTEUx0Npqbe2D9BNCV24G83HTDKzFLlqQWgXCt2FTHLSG8kkNsmI-kh9Sme4FwouQsjh_8HJD6GvlJxSQuRZIaRtaUOIaAhjQ_P0Dm1oy1lDetm_RxvCCWukEL-P0KdS7gmhrRDDR3REhZSc026D_m1L1WPwZTdBrFhHi81OZ20qZP-iq08RJ4dNitavDx3CM_bTlHLVwb-AxRBtqjsIXgdsIAQcfISCXU4TXubLSUucN9jOZl_go1vCU8aPEDU2-lGvFbWcd_3Z9uryF7ZpHgM0NetY_kBcvFnX8hl9cDoU-HK4j9Hd5fb24rq5-Xn14-L8pjF8YLUx2vVWilYKPVLLRgPAGO1AOjpY1wret86OpB_tMLTAOuEkc2YcW80G0oPkx-hkn_uQ098ZSlWTL-tmOkKaixI9Ff0gVuH3N8L7NOfli4pilMuOd5wuIroXmZxKyeDUQ_aTzs-KErVSVHuKaqGoVorqafF8OwTP4wT2v-OAjb8C6VicgA</recordid><startdate>20061201</startdate><enddate>20061201</enddate><creator>Matsuo, Mitsuhiro</creator><creator>Koizumi, Keiichi</creator><creator>Yamada, Sanae</creator><creator>Tomi, Masatoshi</creator><creator>Takahashi, Ri-ichi</creator><creator>Ueda, Masatsugu</creator><creator>Terasaki, Tetsuya</creator><creator>Obinata, Masuo</creator><creator>Hosoya, Ken-ichi</creator><creator>Ohtani, Osamu</creator><creator>Saiki, Ikuo</creator><general>Springer Nature B.V</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>3V.</scope><scope>7QP</scope><scope>7QR</scope><scope>7RV</scope><scope>7SS</scope><scope>7TK</scope><scope>7X7</scope><scope>7XB</scope><scope>88A</scope><scope>88E</scope><scope>8AO</scope><scope>8FD</scope><scope>8FE</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BHPHI</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>KB0</scope><scope>LK8</scope><scope>M0S</scope><scope>M1P</scope><scope>M7P</scope><scope>NAPCQ</scope><scope>P64</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope><scope>RC3</scope><scope>7X8</scope></search><sort><creationdate>20061201</creationdate><title>Establishment and characterization of conditionally immortalized endothelial cell lines from the thoracic duct and inferior vena cava of tsA58/EGFP double-transgenic rats</title><author>Matsuo, Mitsuhiro ; Koizumi, Keiichi ; Yamada, Sanae ; Tomi, Masatoshi ; Takahashi, Ri-ichi ; Ueda, Masatsugu ; Terasaki, Tetsuya ; Obinata, Masuo ; Hosoya, Ken-ichi ; Ohtani, Osamu ; Saiki, Ikuo</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c392t-caf8d76476ab1d2bcee2215e7f19df46384fdb08bd994e256f72fcbb4a2908e73</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2006</creationdate><topic>Animals</topic><topic>Animals, Genetically Modified</topic><topic>Antigens, Viral, Tumor - 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Academic</collection><jtitle>Cell and tissue research</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Matsuo, Mitsuhiro</au><au>Koizumi, Keiichi</au><au>Yamada, Sanae</au><au>Tomi, Masatoshi</au><au>Takahashi, Ri-ichi</au><au>Ueda, Masatsugu</au><au>Terasaki, Tetsuya</au><au>Obinata, Masuo</au><au>Hosoya, Ken-ichi</au><au>Ohtani, Osamu</au><au>Saiki, Ikuo</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Establishment and characterization of conditionally immortalized endothelial cell lines from the thoracic duct and inferior vena cava of tsA58/EGFP double-transgenic rats</atitle><jtitle>Cell and tissue research</jtitle><addtitle>Cell Tissue Res</addtitle><date>2006-12-01</date><risdate>2006</risdate><volume>326</volume><issue>3</issue><spage>749</spage><epage>758</epage><pages>749-758</pages><issn>0302-766X</issn><eissn>1432-0878</eissn><abstract>The basic biology of blood vascular endothelial cells has been well documented. However, little is known about that of lymphatic endothelial cells, despite their importance under normal and pathological conditions. The lack of a lymphatic endothelial cell line has hampered progress in this field. The objective of this study has been to establish and characterize lymphatic and venous endothelial cell lines derived from newly developed tsA58/EGFP transgenic rats harboring the temperature-sensitive simian virus 40 (SV40) large T-antigen and enhanced green fluorescent protein (EGFP). Endothelial cells were isolated from the transgenic rats by intraluminal enzymatic digestion. The cloned cell lines were named TR-LE (temperature-sensitive rat lymphatic endothelial cells from thoracic duct) and TR-BE (temperature-sensitive rat blood-vessel endothelial cells from inferior vena cava), respectively, and cultured on fibronectin-coated dishes in HuMedia-EG2 supplemented with 20% fetal bovine serum and Endothelial Mitogen at a permissive temperature, 33 degrees C. A temperature shift to 37 degrees C resulted in a decrease in proliferation with degradation of the large T-antigen and cleavage of poly (ADP-ribose) polymerase. TR-LE cells expressed lymphatic endothelial markers VEGFR-3 (vascular endothelial growth factor receptor), LYVE-1 (a lymphatic endothelial receptor), Prox-1 (a homeobox gene product), and podoplanin (a glomerular podocyte membrane mucoprotein), together with endothelial markers CD31, Tie-2, and VEGFR-2, whereas TR-BE cells expressed CD31, Tie-2, and VEGFR-2, but no lymphatic endothelial markers. Thus, these conditionally immortalized and EGFP-expressing lymphatic and vascular endothelial cell lines might represent an important tool for the study of endothelial cell functions in vitro.</abstract><cop>Germany</cop><pub>Springer Nature B.V</pub><pmid>16773315</pmid><doi>10.1007/s00441-006-0229-x</doi><tpages>10</tpages></addata></record> |
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| subjects | Animals Animals, Genetically Modified Antigens, Viral, Tumor - genetics Biomarkers - analysis Cell Culture Techniques Cell Line, Transformed Cell Proliferation Cell Transformation, Viral Cells Clone Cells Endothelial Cells - cytology Endothelial Cells - metabolism Endothelial Cells - physiology Endothelial Cells - ultrastructure Endothelium, Lymphatic - cytology Endothelium, Vascular - cytology Enzymes Fluorescent Dyes - metabolism Gene Expression Profiling Green Fluorescent Proteins - metabolism Lymphatic system Pulmonary arteries Rats Rodents Simian virus 40 - physiology Temperature Thoracic Duct - cytology Vena Cava, Inferior - cytology |
| title | Establishment and characterization of conditionally immortalized endothelial cell lines from the thoracic duct and inferior vena cava of tsA58/EGFP double-transgenic rats |
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