Transformation of rat inner medullary fibroblasts to myofibroblasts in vitro
Transformation of rat inner medullary fibroblasts to myofibroblasts in vitro. Renal fibroblasts play a major role in the pathogenesis of renal interstitial fibrosis. This process is associated at least in some forms of interstitial fibrosis with a differentiation of fibroblasts into myofibroblasts,...
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Veröffentlicht in: | Kidney international 1997-11, Vol.52 (5), p.1279-1290 |
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description | Transformation of rat inner medullary fibroblasts to myofibroblasts in vitro. Renal fibroblasts play a major role in the pathogenesis of renal interstitial fibrosis. This process is associated at least in some forms of interstitial fibrosis with a differentiation of fibroblasts into myofibroblasts, characterized by the de novo expression of α-smooth muscle (α-sm) actin and/or desmin. Both the mechanisms underlying this differentiation and their effects on cellular function are poorly understood. In vitro studies are difficult since the phenotypes of fibroblasts in culture have as yet not been well defined. We have, therefore, examined the phenotype of inner medullary fibroblasts (IMF) during the transition from in vivo to in vitro in various cell fractions derived from the inner medulla of healthy rats. IMF were positive for the lectin BSL-1 and negative for markers of endothelial cells. IMF first lost their prominent lipid droplets in vitro. Subsequently they developed cytoplasmic processes accompanied by a decrease in their reactivity for the lectin BSL-1 from strong to weak. From day 3 in primary culture, exclusively these weakly positive BSL-1 cells showed a de novo expression of α-sm actin (day 4 of primary culture, 75 ± 4% day 20, 94 ± 2%) and desmin (day 4, 43 ± 8% day 20, 66 ± 6%), classifying them as myofibroblasts. This transformation depended on culture conditions. In a mixed coculture with inner medullary collecting duct (IMCD) cells the transformation of IMF was largely absent: a significantly greater number of strong BSL-1 positive cells contained prominent lipid droplets (39 ± 4 vs. 19 ± 4%, P < 0.05) on day 4 of primary culture, and the transition of strongly to weakly positive BSL-1 IMF was almost completely blocked. By reducing the seeding density of IMCD cells the effect of this condition on IMF transformation could be largely abolished. This first detailed pheno-typic characterization of rat fibroblasts during the transition from in vivo to in vitro demonstrates that these cells-depending on culture conditions-differentiate to myofibroblasts within a few days of primary culture and that subcultured IMF exhibit predominantly this phenotype. The presented model may serve as a useful tool for the in vitro study of myofibroblast formation and the consequences of such a differentiation for the physiological functions of IMF. |
doi_str_mv | 10.1038/ki.1997.453 |
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Renal fibroblasts play a major role in the pathogenesis of renal interstitial fibrosis. This process is associated at least in some forms of interstitial fibrosis with a differentiation of fibroblasts into myofibroblasts, characterized by the de novo expression of α-smooth muscle (α-sm) actin and/or desmin. Both the mechanisms underlying this differentiation and their effects on cellular function are poorly understood. In vitro studies are difficult since the phenotypes of fibroblasts in culture have as yet not been well defined. We have, therefore, examined the phenotype of inner medullary fibroblasts (IMF) during the transition from in vivo to in vitro in various cell fractions derived from the inner medulla of healthy rats. IMF were positive for the lectin BSL-1 and negative for markers of endothelial cells. IMF first lost their prominent lipid droplets in vitro. Subsequently they developed cytoplasmic processes accompanied by a decrease in their reactivity for the lectin BSL-1 from strong to weak. From day 3 in primary culture, exclusively these weakly positive BSL-1 cells showed a de novo expression of α-sm actin (day 4 of primary culture, 75 ± 4% day 20, 94 ± 2%) and desmin (day 4, 43 ± 8% day 20, 66 ± 6%), classifying them as myofibroblasts. This transformation depended on culture conditions. In a mixed coculture with inner medullary collecting duct (IMCD) cells the transformation of IMF was largely absent: a significantly greater number of strong BSL-1 positive cells contained prominent lipid droplets (39 ± 4 vs. 19 ± 4%, P < 0.05) on day 4 of primary culture, and the transition of strongly to weakly positive BSL-1 IMF was almost completely blocked. By reducing the seeding density of IMCD cells the effect of this condition on IMF transformation could be largely abolished. This first detailed pheno-typic characterization of rat fibroblasts during the transition from in vivo to in vitro demonstrates that these cells-depending on culture conditions-differentiate to myofibroblasts within a few days of primary culture and that subcultured IMF exhibit predominantly this phenotype. The presented model may serve as a useful tool for the in vitro study of myofibroblast formation and the consequences of such a differentiation for the physiological functions of IMF.</description><identifier>ISSN: 0085-2538</identifier><identifier>EISSN: 1523-1755</identifier><identifier>DOI: 10.1038/ki.1997.453</identifier><identifier>PMID: 9350651</identifier><identifier>CODEN: KDYIA5</identifier><language>eng</language><publisher>New York, NY: Elsevier Inc</publisher><subject>Actins - analysis ; Animals ; bandeiraea simplifolia-1 lectin ; Biological and medical sciences ; Cell Differentiation ; cell isolation ; Cells, Cultured ; desmin ; Desmin - analysis ; Dinoprostone - biosynthesis ; Fibroblasts - cytology ; Immunohistochemistry ; inner medulla ; Interstitial nephritis ; Kidney Medulla - chemistry ; Kidney Medulla - cytology ; lipid laden cells ; Male ; Medical sciences ; myofibroblast ; Nephrology. Urinary tract diseases ; Nephropathies. Renovascular diseases. Renal failure ; Phenotype ; primary culture ; Rats ; Rats, Wistar ; renal fibroblasts ; α-smooth muscle actin</subject><ispartof>Kidney international, 1997-11, Vol.52 (5), p.1279-1290</ispartof><rights>1997 International Society of Nephrology</rights><rights>1998 INIST-CNRS</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c462t-72859c30127625d3234a8df0b4003a7288883e4f0d0f4596f0b55822f041a42a3</citedby><cites>FETCH-LOGICAL-c462t-72859c30127625d3234a8df0b4003a7288883e4f0d0f4596f0b55822f041a42a3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>309,310,314,780,784,789,790,23930,23931,25140,27924,27925</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=2050301$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/9350651$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Grupp, Clemens</creatorcontrib><creatorcontrib>Lottermoser, Jens</creatorcontrib><creatorcontrib>Cohen, David I.</creatorcontrib><creatorcontrib>Begher, Michael</creatorcontrib><creatorcontrib>Franz, Hans-Eduard</creatorcontrib><creatorcontrib>Müller, Gerhard A.</creatorcontrib><title>Transformation of rat inner medullary fibroblasts to myofibroblasts in vitro</title><title>Kidney international</title><addtitle>Kidney Int</addtitle><description>Transformation of rat inner medullary fibroblasts to myofibroblasts in vitro. Renal fibroblasts play a major role in the pathogenesis of renal interstitial fibrosis. This process is associated at least in some forms of interstitial fibrosis with a differentiation of fibroblasts into myofibroblasts, characterized by the de novo expression of α-smooth muscle (α-sm) actin and/or desmin. Both the mechanisms underlying this differentiation and their effects on cellular function are poorly understood. In vitro studies are difficult since the phenotypes of fibroblasts in culture have as yet not been well defined. We have, therefore, examined the phenotype of inner medullary fibroblasts (IMF) during the transition from in vivo to in vitro in various cell fractions derived from the inner medulla of healthy rats. IMF were positive for the lectin BSL-1 and negative for markers of endothelial cells. IMF first lost their prominent lipid droplets in vitro. Subsequently they developed cytoplasmic processes accompanied by a decrease in their reactivity for the lectin BSL-1 from strong to weak. From day 3 in primary culture, exclusively these weakly positive BSL-1 cells showed a de novo expression of α-sm actin (day 4 of primary culture, 75 ± 4% day 20, 94 ± 2%) and desmin (day 4, 43 ± 8% day 20, 66 ± 6%), classifying them as myofibroblasts. This transformation depended on culture conditions. In a mixed coculture with inner medullary collecting duct (IMCD) cells the transformation of IMF was largely absent: a significantly greater number of strong BSL-1 positive cells contained prominent lipid droplets (39 ± 4 vs. 19 ± 4%, P < 0.05) on day 4 of primary culture, and the transition of strongly to weakly positive BSL-1 IMF was almost completely blocked. By reducing the seeding density of IMCD cells the effect of this condition on IMF transformation could be largely abolished. This first detailed pheno-typic characterization of rat fibroblasts during the transition from in vivo to in vitro demonstrates that these cells-depending on culture conditions-differentiate to myofibroblasts within a few days of primary culture and that subcultured IMF exhibit predominantly this phenotype. The presented model may serve as a useful tool for the in vitro study of myofibroblast formation and the consequences of such a differentiation for the physiological functions of IMF.</description><subject>Actins - analysis</subject><subject>Animals</subject><subject>bandeiraea simplifolia-1 lectin</subject><subject>Biological and medical sciences</subject><subject>Cell Differentiation</subject><subject>cell isolation</subject><subject>Cells, Cultured</subject><subject>desmin</subject><subject>Desmin - analysis</subject><subject>Dinoprostone - biosynthesis</subject><subject>Fibroblasts - cytology</subject><subject>Immunohistochemistry</subject><subject>inner medulla</subject><subject>Interstitial nephritis</subject><subject>Kidney Medulla - chemistry</subject><subject>Kidney Medulla - cytology</subject><subject>lipid laden cells</subject><subject>Male</subject><subject>Medical sciences</subject><subject>myofibroblast</subject><subject>Nephrology. Urinary tract diseases</subject><subject>Nephropathies. Renovascular diseases. Renal failure</subject><subject>Phenotype</subject><subject>primary culture</subject><subject>Rats</subject><subject>Rats, Wistar</subject><subject>renal fibroblasts</subject><subject>α-smooth muscle actin</subject><issn>0085-2538</issn><issn>1523-1755</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1997</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNptkEtrAyEURqW0pGnaVdcFF6WbMulVx3ksS-gLAt2ka3EcBZuZMVUTyL-vIUPoom5Ev8PV7yB0S2BOgFVPazsndV3Oc87O0JRwyjJScn6OpgAVzyhn1SW6CuEb0rlmMEGTmnEoOJmi5crLIRjnexmtG7Az2MuI7TBoj3vdbrtO-j02tvGu6WSIAUeH-737e2MHvLPRu2t0YWQX9M24z9DX68tq8Z4tP98-Fs_LTOUFjVlJK14rBoSWBeUtoyyXVWugyQGYTGlaTOcGWjA5r4uUcF5RaiAnMqeSzdDDce7Gu5-tDlH0Niidvjpotw2irFkJnNIEPh5B5V0IXhux8bZPhQQBcXAn1lYc3InkLtF349htk6qf2FFWyu_HXAYlO5PMKRtOGAUOqVTC-BHTScHOai-CsnpQurVeqyhaZ_99_hea_Ieo</recordid><startdate>19971101</startdate><enddate>19971101</enddate><creator>Grupp, Clemens</creator><creator>Lottermoser, Jens</creator><creator>Cohen, David I.</creator><creator>Begher, Michael</creator><creator>Franz, Hans-Eduard</creator><creator>Müller, Gerhard A.</creator><general>Elsevier Inc</general><general>Nature Publishing</general><scope>6I.</scope><scope>AAFTH</scope><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>7X8</scope></search><sort><creationdate>19971101</creationdate><title>Transformation of rat inner medullary fibroblasts to myofibroblasts in vitro</title><author>Grupp, Clemens ; Lottermoser, Jens ; Cohen, David I. ; Begher, Michael ; Franz, Hans-Eduard ; Müller, Gerhard A.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c462t-72859c30127625d3234a8df0b4003a7288883e4f0d0f4596f0b55822f041a42a3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1997</creationdate><topic>Actins - analysis</topic><topic>Animals</topic><topic>bandeiraea simplifolia-1 lectin</topic><topic>Biological and medical sciences</topic><topic>Cell Differentiation</topic><topic>cell isolation</topic><topic>Cells, Cultured</topic><topic>desmin</topic><topic>Desmin - analysis</topic><topic>Dinoprostone - biosynthesis</topic><topic>Fibroblasts - cytology</topic><topic>Immunohistochemistry</topic><topic>inner medulla</topic><topic>Interstitial nephritis</topic><topic>Kidney Medulla - chemistry</topic><topic>Kidney Medulla - cytology</topic><topic>lipid laden cells</topic><topic>Male</topic><topic>Medical sciences</topic><topic>myofibroblast</topic><topic>Nephrology. Urinary tract diseases</topic><topic>Nephropathies. Renovascular diseases. Renal failure</topic><topic>Phenotype</topic><topic>primary culture</topic><topic>Rats</topic><topic>Rats, Wistar</topic><topic>renal fibroblasts</topic><topic>α-smooth muscle actin</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Grupp, Clemens</creatorcontrib><creatorcontrib>Lottermoser, Jens</creatorcontrib><creatorcontrib>Cohen, David I.</creatorcontrib><creatorcontrib>Begher, Michael</creatorcontrib><creatorcontrib>Franz, Hans-Eduard</creatorcontrib><creatorcontrib>Müller, Gerhard A.</creatorcontrib><collection>ScienceDirect Open Access Titles</collection><collection>Elsevier:ScienceDirect:Open Access</collection><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>MEDLINE - Academic</collection><jtitle>Kidney international</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Grupp, Clemens</au><au>Lottermoser, Jens</au><au>Cohen, David I.</au><au>Begher, Michael</au><au>Franz, Hans-Eduard</au><au>Müller, Gerhard A.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Transformation of rat inner medullary fibroblasts to myofibroblasts in vitro</atitle><jtitle>Kidney international</jtitle><addtitle>Kidney Int</addtitle><date>1997-11-01</date><risdate>1997</risdate><volume>52</volume><issue>5</issue><spage>1279</spage><epage>1290</epage><pages>1279-1290</pages><issn>0085-2538</issn><eissn>1523-1755</eissn><coden>KDYIA5</coden><abstract>Transformation of rat inner medullary fibroblasts to myofibroblasts in vitro. Renal fibroblasts play a major role in the pathogenesis of renal interstitial fibrosis. This process is associated at least in some forms of interstitial fibrosis with a differentiation of fibroblasts into myofibroblasts, characterized by the de novo expression of α-smooth muscle (α-sm) actin and/or desmin. Both the mechanisms underlying this differentiation and their effects on cellular function are poorly understood. In vitro studies are difficult since the phenotypes of fibroblasts in culture have as yet not been well defined. We have, therefore, examined the phenotype of inner medullary fibroblasts (IMF) during the transition from in vivo to in vitro in various cell fractions derived from the inner medulla of healthy rats. IMF were positive for the lectin BSL-1 and negative for markers of endothelial cells. IMF first lost their prominent lipid droplets in vitro. Subsequently they developed cytoplasmic processes accompanied by a decrease in their reactivity for the lectin BSL-1 from strong to weak. From day 3 in primary culture, exclusively these weakly positive BSL-1 cells showed a de novo expression of α-sm actin (day 4 of primary culture, 75 ± 4% day 20, 94 ± 2%) and desmin (day 4, 43 ± 8% day 20, 66 ± 6%), classifying them as myofibroblasts. This transformation depended on culture conditions. In a mixed coculture with inner medullary collecting duct (IMCD) cells the transformation of IMF was largely absent: a significantly greater number of strong BSL-1 positive cells contained prominent lipid droplets (39 ± 4 vs. 19 ± 4%, P < 0.05) on day 4 of primary culture, and the transition of strongly to weakly positive BSL-1 IMF was almost completely blocked. By reducing the seeding density of IMCD cells the effect of this condition on IMF transformation could be largely abolished. This first detailed pheno-typic characterization of rat fibroblasts during the transition from in vivo to in vitro demonstrates that these cells-depending on culture conditions-differentiate to myofibroblasts within a few days of primary culture and that subcultured IMF exhibit predominantly this phenotype. The presented model may serve as a useful tool for the in vitro study of myofibroblast formation and the consequences of such a differentiation for the physiological functions of IMF.</abstract><cop>New York, NY</cop><pub>Elsevier Inc</pub><pmid>9350651</pmid><doi>10.1038/ki.1997.453</doi><tpages>12</tpages><oa>free_for_read</oa></addata></record> |
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subjects | Actins - analysis Animals bandeiraea simplifolia-1 lectin Biological and medical sciences Cell Differentiation cell isolation Cells, Cultured desmin Desmin - analysis Dinoprostone - biosynthesis Fibroblasts - cytology Immunohistochemistry inner medulla Interstitial nephritis Kidney Medulla - chemistry Kidney Medulla - cytology lipid laden cells Male Medical sciences myofibroblast Nephrology. Urinary tract diseases Nephropathies. Renovascular diseases. Renal failure Phenotype primary culture Rats Rats, Wistar renal fibroblasts α-smooth muscle actin |
title | Transformation of rat inner medullary fibroblasts to myofibroblasts in vitro |
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