Endothelial Cell Modulation of Smooth Muscle Cell Morphology and Organizational Growth Pattern
Intimal hyperplasia is characterized by smooth muscle cell (SMC) dedifferentiation from a contractile to a synthetic phenotype prior to migration and proliferation. Regulatory mechanisms controlling SMC phenotype are not well known. This study examined the effect of endothelial cells (ECs) on SMC mo...
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| Veröffentlicht in: | Annals of vascular surgery 1996, Vol.10 (1), p.4-10 |
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| creator | Powell, Richard J. Cronenwett, Jack L. Fillinger, Mark F. Wagner, Robert J. Sampson, Lawrence N. |
| description | Intimal hyperplasia is characterized by smooth muscle cell (SMC) dedifferentiation from a contractile to a synthetic phenotype prior to migration and proliferation. Regulatory mechanisms controlling SMC phenotype are not well known. This study examined the effect of endothelial cells (ECs) on SMC morphology in coculture. Subcultured bovine ECs and SMCs were plated on opposite sides of a 13 μm thick, semipermeable membrane (0.45 μm pores, Cyclopore) to allow potential humoral and cellular cross-membrane communication. SMCs were studied (5 wells/group) in coculture opposite confluent ECs (EC/SMC) and alone (SMC controls). After 4 days of culture in Dulbecco's modified Eagle medium/2.5% calf serum, SMCs were harvested. The ratio of protein/DNA was measured as an index of SMC hypertrophy (synthetic SMC phenotype). SMCs were examined with light and scanning electron microscopy to evaluate cell surface area, cellular morphology, and macroscopic growth characteristics. Flow cytometry was used to determine the cellular RNA/DNA ratio. SMC control cultures had a significantly greater protein-to-DNA content than SMCs cocultured with ECs (175 ± 9 vs. 115 ± 7 μg protein/μg DMA;
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| doi_str_mv | 10.1007/BF02002334 |
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p <0.001). SMC control cultures also had 6.5 times greater cell surface area (5.8 ± 0.3 × 10
3 μm
2) than cocultured SMCs (0.9 ± 0.1;
p <0.001). In SMC control cultures, SMC hypertrophy and rapid “hill and valley” formation were observed. In contrast, SMCs from the EC/SMC group exhibited a more spindle-shaped, contractile-appearing phenotype with more uniform, evenly distributed cells and no hill and valley formation. SMC control cultures also had a higher RNA/DNA ratio. Thus the presence of confluent ECs substantially altered the morphology and growth characteristics normally observed for SMCs in vitro. This coculture system provides a model to further study EC-SMC interaction, which could have important in vivo consequences.</description><identifier>ISSN: 0890-5096</identifier><identifier>EISSN: 1615-5947</identifier><identifier>DOI: 10.1007/BF02002334</identifier><identifier>PMID: 8688295</identifier><language>eng</language><publisher>Netherlands: Elsevier Inc</publisher><subject>Animals ; Cattle ; Cell Differentiation ; Cell Division ; Cells, Cultured ; DNA - analysis ; Endothelium, Vascular - cytology ; Endothelium, Vascular - ultrastructure ; Flow Cytometry ; Fluorometry ; Muscle Contraction ; Muscle, Smooth, Vascular - cytology ; Muscle, Smooth, Vascular - metabolism ; Muscle, Smooth, Vascular - ultrastructure ; Phenotype ; RNA - analysis</subject><ispartof>Annals of vascular surgery, 1996, Vol.10 (1), p.4-10</ispartof><rights>1996 Annals of Vascular Surgery, Inc.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c327t-1b19344dfbec4080db174a45d553ff1f40e485bd05346a93138ea9e7c8a5ef653</citedby><cites>FETCH-LOGICAL-c327t-1b19344dfbec4080db174a45d553ff1f40e485bd05346a93138ea9e7c8a5ef653</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1007/BF02002334$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,780,784,3548,4021,27921,27922,27923,45993</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/8688295$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Powell, Richard J.</creatorcontrib><creatorcontrib>Cronenwett, Jack L.</creatorcontrib><creatorcontrib>Fillinger, Mark F.</creatorcontrib><creatorcontrib>Wagner, Robert J.</creatorcontrib><creatorcontrib>Sampson, Lawrence N.</creatorcontrib><title>Endothelial Cell Modulation of Smooth Muscle Cell Morphology and Organizational Growth Pattern</title><title>Annals of vascular surgery</title><addtitle>Ann Vasc Surg</addtitle><description>Intimal hyperplasia is characterized by smooth muscle cell (SMC) dedifferentiation from a contractile to a synthetic phenotype prior to migration and proliferation. Regulatory mechanisms controlling SMC phenotype are not well known. This study examined the effect of endothelial cells (ECs) on SMC morphology in coculture. Subcultured bovine ECs and SMCs were plated on opposite sides of a 13 μm thick, semipermeable membrane (0.45 μm pores, Cyclopore) to allow potential humoral and cellular cross-membrane communication. SMCs were studied (5 wells/group) in coculture opposite confluent ECs (EC/SMC) and alone (SMC controls). After 4 days of culture in Dulbecco's modified Eagle medium/2.5% calf serum, SMCs were harvested. The ratio of protein/DNA was measured as an index of SMC hypertrophy (synthetic SMC phenotype). SMCs were examined with light and scanning electron microscopy to evaluate cell surface area, cellular morphology, and macroscopic growth characteristics. Flow cytometry was used to determine the cellular RNA/DNA ratio. SMC control cultures had a significantly greater protein-to-DNA content than SMCs cocultured with ECs (175 ± 9 vs. 115 ± 7 μg protein/μg DMA;
p <0.001). SMC control cultures also had 6.5 times greater cell surface area (5.8 ± 0.3 × 10
3 μm
2) than cocultured SMCs (0.9 ± 0.1;
p <0.001). In SMC control cultures, SMC hypertrophy and rapid “hill and valley” formation were observed. In contrast, SMCs from the EC/SMC group exhibited a more spindle-shaped, contractile-appearing phenotype with more uniform, evenly distributed cells and no hill and valley formation. SMC control cultures also had a higher RNA/DNA ratio. Thus the presence of confluent ECs substantially altered the morphology and growth characteristics normally observed for SMCs in vitro. This coculture system provides a model to further study EC-SMC interaction, which could have important in vivo consequences.</description><subject>Animals</subject><subject>Cattle</subject><subject>Cell Differentiation</subject><subject>Cell Division</subject><subject>Cells, Cultured</subject><subject>DNA - analysis</subject><subject>Endothelium, Vascular - cytology</subject><subject>Endothelium, Vascular - ultrastructure</subject><subject>Flow Cytometry</subject><subject>Fluorometry</subject><subject>Muscle Contraction</subject><subject>Muscle, Smooth, Vascular - cytology</subject><subject>Muscle, Smooth, Vascular - metabolism</subject><subject>Muscle, Smooth, Vascular - ultrastructure</subject><subject>Phenotype</subject><subject>RNA - analysis</subject><issn>0890-5096</issn><issn>1615-5947</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1996</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNptkDFPwzAQRi0EKqWwsCNlYkAK2LGdOCNUbUFqVSRgJXLsS2uUxsVOQOXXY2iBhemG792nu4fQKcGXBOPs6maME4wTStke6pOU8JjnLNtHfSxyHHOcp4foyPsXjEkimOihnkiFSHLeR8-jRtt2CbWRdTSEuo5mVne1bI1tIltFDysb4mjWeVXDD-DWS1vbxSaSjY7mbiEb8_G9ETomzr6HhXvZtuCaY3RQydrDyW4O0NN49Di8jafzyd3wehormmRtTEqSU8Z0VYJiWGBdkoxJxjXntKpIxTAwwUuNOWWpzCmhAmQOmRKSQ5VyOkDn2961s68d-LZYGa_CtbIB2_kiE0EUT1kAL7agctZ7B1WxdmYl3aYguPiSWfzJDPDZrrUrV6B_0Z29kLNtDuG1NwOu8MpAo0AbB6ottDX_1X4CTYGAKQ</recordid><startdate>1996</startdate><enddate>1996</enddate><creator>Powell, Richard J.</creator><creator>Cronenwett, Jack L.</creator><creator>Fillinger, Mark F.</creator><creator>Wagner, Robert J.</creator><creator>Sampson, Lawrence N.</creator><general>Elsevier Inc</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>7X8</scope></search><sort><creationdate>1996</creationdate><title>Endothelial Cell Modulation of Smooth Muscle Cell Morphology and Organizational Growth Pattern</title><author>Powell, Richard J. ; Cronenwett, Jack L. ; Fillinger, Mark F. ; Wagner, Robert J. ; Sampson, Lawrence N.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c327t-1b19344dfbec4080db174a45d553ff1f40e485bd05346a93138ea9e7c8a5ef653</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1996</creationdate><topic>Animals</topic><topic>Cattle</topic><topic>Cell Differentiation</topic><topic>Cell Division</topic><topic>Cells, Cultured</topic><topic>DNA - analysis</topic><topic>Endothelium, Vascular - cytology</topic><topic>Endothelium, Vascular - ultrastructure</topic><topic>Flow Cytometry</topic><topic>Fluorometry</topic><topic>Muscle Contraction</topic><topic>Muscle, Smooth, Vascular - cytology</topic><topic>Muscle, Smooth, Vascular - metabolism</topic><topic>Muscle, Smooth, Vascular - ultrastructure</topic><topic>Phenotype</topic><topic>RNA - analysis</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Powell, Richard J.</creatorcontrib><creatorcontrib>Cronenwett, Jack L.</creatorcontrib><creatorcontrib>Fillinger, Mark F.</creatorcontrib><creatorcontrib>Wagner, Robert J.</creatorcontrib><creatorcontrib>Sampson, Lawrence N.</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>Annals of vascular surgery</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Powell, Richard J.</au><au>Cronenwett, Jack L.</au><au>Fillinger, Mark F.</au><au>Wagner, Robert J.</au><au>Sampson, Lawrence N.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Endothelial Cell Modulation of Smooth Muscle Cell Morphology and Organizational Growth Pattern</atitle><jtitle>Annals of vascular surgery</jtitle><addtitle>Ann Vasc Surg</addtitle><date>1996</date><risdate>1996</risdate><volume>10</volume><issue>1</issue><spage>4</spage><epage>10</epage><pages>4-10</pages><issn>0890-5096</issn><eissn>1615-5947</eissn><abstract>Intimal hyperplasia is characterized by smooth muscle cell (SMC) dedifferentiation from a contractile to a synthetic phenotype prior to migration and proliferation. Regulatory mechanisms controlling SMC phenotype are not well known. This study examined the effect of endothelial cells (ECs) on SMC morphology in coculture. Subcultured bovine ECs and SMCs were plated on opposite sides of a 13 μm thick, semipermeable membrane (0.45 μm pores, Cyclopore) to allow potential humoral and cellular cross-membrane communication. SMCs were studied (5 wells/group) in coculture opposite confluent ECs (EC/SMC) and alone (SMC controls). After 4 days of culture in Dulbecco's modified Eagle medium/2.5% calf serum, SMCs were harvested. The ratio of protein/DNA was measured as an index of SMC hypertrophy (synthetic SMC phenotype). SMCs were examined with light and scanning electron microscopy to evaluate cell surface area, cellular morphology, and macroscopic growth characteristics. Flow cytometry was used to determine the cellular RNA/DNA ratio. SMC control cultures had a significantly greater protein-to-DNA content than SMCs cocultured with ECs (175 ± 9 vs. 115 ± 7 μg protein/μg DMA;
p <0.001). SMC control cultures also had 6.5 times greater cell surface area (5.8 ± 0.3 × 10
3 μm
2) than cocultured SMCs (0.9 ± 0.1;
p <0.001). In SMC control cultures, SMC hypertrophy and rapid “hill and valley” formation were observed. In contrast, SMCs from the EC/SMC group exhibited a more spindle-shaped, contractile-appearing phenotype with more uniform, evenly distributed cells and no hill and valley formation. SMC control cultures also had a higher RNA/DNA ratio. Thus the presence of confluent ECs substantially altered the morphology and growth characteristics normally observed for SMCs in vitro. This coculture system provides a model to further study EC-SMC interaction, which could have important in vivo consequences.</abstract><cop>Netherlands</cop><pub>Elsevier Inc</pub><pmid>8688295</pmid><doi>10.1007/BF02002334</doi><tpages>7</tpages></addata></record> |
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| ispartof | Annals of vascular surgery, 1996, Vol.10 (1), p.4-10 |
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| subjects | Animals Cattle Cell Differentiation Cell Division Cells, Cultured DNA - analysis Endothelium, Vascular - cytology Endothelium, Vascular - ultrastructure Flow Cytometry Fluorometry Muscle Contraction Muscle, Smooth, Vascular - cytology Muscle, Smooth, Vascular - metabolism Muscle, Smooth, Vascular - ultrastructure Phenotype RNA - analysis |
| title | Endothelial Cell Modulation of Smooth Muscle Cell Morphology and Organizational Growth Pattern |
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