Cytoskeletal organization of human mesenchymal stem cells (MSC) changes during their osteogenic differentiation
Identifieur interne : 007636 ( Istex/Corpus ); précédent : 007635; suivant : 007637Cytoskeletal organization of human mesenchymal stem cells (MSC) changes during their osteogenic differentiation
Auteurs : J. Pablo Rodríguez ; Mauricio González ; Susana Ríos ; Ver Nica CambiazoSource :
- Journal of Cellular Biochemistry [ 0730-2312 ] ; 2004-11-01.
English descriptors
- KwdEn :
- Actin, Actin cytoskeleton, Actin organization, Alexa fluor, Alkaline, Alkaline phosphatase activity, Basal, Basal conditions, Biochem, Biochemical markers, Bone marrow, Calcium deposition, Calcium phosphate deposition, Cell biochem, Cell biochem koukouritaki, Cell layer, Cell morphology, Cell shape, Characteristic phenotype, Chile, Cortical organization, Culture medium, Cytochalasin, Cytoskeletal, Cytoskeletal integrity, Cytoskeletal organization, Cytoskeleton, Cytoskeleton organization, Different donors, Differentiation, Differentiation markers, Entire cytoplasm, Human mesenchymal, Human mscs, Koukouritaki, Mesenchymal, Microtubule, Microtubule network, Morphological changes, Morphology, Msc, Nocodazole, Normal bone, Osteoblast, Osteoblastic, Osteogenic, Osteogenic conditions, Osteogenic differentiation, Osteogenic medium, Osteoporotic, Phenotype, Phosphatase, Rapid changes, Results show, Tubulin, Untreated, Untreated cells, Upper panels.
- Teeft :
- Actin, Actin cytoskeleton, Actin organization, Alexa fluor, Alkaline, Alkaline phosphatase activity, Basal, Basal conditions, Biochem, Biochemical markers, Bone marrow, Calcium deposition, Calcium phosphate deposition, Cell biochem, Cell biochem koukouritaki, Cell layer, Cell morphology, Cell shape, Characteristic phenotype, Chile, Cortical organization, Culture medium, Cytochalasin, Cytoskeletal, Cytoskeletal integrity, Cytoskeletal organization, Cytoskeleton, Cytoskeleton organization, Different donors, Differentiation, Differentiation markers, Entire cytoplasm, Human mesenchymal, Human mscs, Koukouritaki, Mesenchymal, Microtubule, Microtubule network, Morphological changes, Morphology, Msc, Nocodazole, Normal bone, Osteoblast, Osteoblastic, Osteogenic, Osteogenic conditions, Osteogenic differentiation, Osteogenic medium, Osteoporotic, Phenotype, Phosphatase, Rapid changes, Results show, Tubulin, Untreated, Untreated cells, Upper panels.
Abstract
Human MSCs have been studied to define the mechanisms involved in normal bone remodeling and the regulation of osteogenesis. During osteogenic differentiation, MSCs change from their characteristic fibroblast‐like phenotype to near spherical shape. In this study, we analyzed the correlation between the organization of cytoskeleton of MSCs, changes in cell morphology, and the expression of specific markers (alkaline phosphatase activity and calcium deposition) of osteogenic differentiation. For osteoblastic differentiation, cells were cultured in a culture medium supplemented with 100 nM dexamethasone, 10 mM β‐ glycerophosphate, and 50 μg/ml ascorbic acid. The organization of microfilaments and microtubules was examined by inmunofluorescence using Alexa fluor 594 phalloidin and anti α‐tubulin monoclonal antibody. Cytochalasin D and nocodazole were used to alter reversibly the cytoskeleton dynamic. A remarkable change in cytoskeleton organization was observed in human MSCs during osteogenic differentiation. Actin cytoskeleton changed from a large number of thin, parallel microfilament bundles extending across the entire cytoplasm in undifferentiated MSCs to a few thick actin filament bundles located at the outermost periphery in differentiated cells. Under osteogenic culture conditions, a reversible reorganization of microfilaments induced by an initial treatment with cytochalasin D but not with nocodazole reduced the expression of differentiation markers, without affecting the final morphology of the cells. The results indicate that changes in the assembly and disassembly kinetics of microfilaments dynamic of actin network formation may be critical in supporting the osteogenic differentiation of human MSCs; also indicated that the organization of microtubules appears to have a regulatory role on the kinetic of this process. © 2004 Wiley‐Liss, Inc.
Url:
DOI: 10.1002/jcb.20234
Links to Exploration step
ISTEX:EEAEC74F03B67EADDC845D9F00ABA023094960BBLe document en format XML
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Pablo Rodríguez</name><affiliation><mods:affiliation>Laboratorio de Biología Celular, INTA, Universidad de Chile, Casilla 138‐11, Santiago, Chile</mods:affiliation></affiliation><affiliation><mods:affiliation>E-mail: jprodrig@inta.cl</mods:affiliation></affiliation><affiliation><mods:affiliation>Correspondence address: Laboratorio de Biología Celular, INTA, Universidad de Chile, Casilla 138‐11, Santiago, Chile.</mods:affiliation></affiliation></author><author><name sortKey="Gonzalez, Mauricio" sort="Gonzalez, Mauricio" uniqKey="Gonzalez M" first="Mauricio" last="González">Mauricio González</name><affiliation><mods:affiliation>Laboratorio de Bioinformática y Expresión Génica, INTA, Universidad de Chile, Casilla 138‐11, Santiago, Chile</mods:affiliation></affiliation></author><author><name sortKey="Rios, Susana" sort="Rios, Susana" uniqKey="Rios S" first="Susana" last="Ríos">Susana Ríos</name><affiliation><mods:affiliation>Laboratorio de Biología Celular, INTA, Universidad de Chile, Casilla 138‐11, Santiago, Chile</mods:affiliation></affiliation></author><author><name sortKey="Cambiazo, Ver Nica" sort="Cambiazo, Ver Nica" uniqKey="Cambiazo V" first="Ver Nica" last="Cambiazo">Ver Nica Cambiazo</name><affiliation><mods:affiliation>Laboratorio de Bioinformática y Expresión Génica, INTA, Universidad de Chile, Casilla 138‐11, Santiago, Chile</mods:affiliation></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">ISTEX</idno><idno type="RBID">ISTEX:EEAEC74F03B67EADDC845D9F00ABA023094960BB</idno><date when="2004" year="2004">2004</date><idno type="doi">10.1002/jcb.20234</idno><idno type="url">https://api.istex.fr/document/EEAEC74F03B67EADDC845D9F00ABA023094960BB/fulltext/pdf</idno><idno type="wicri:Area/Istex/Corpus">007636</idno><idno type="wicri:explorRef" wicri:stream="Istex" wicri:step="Corpus" wicri:corpus="ISTEX">007636</idno></publicationStmt><sourceDesc><biblStruct><analytic><title level="a" type="main" xml:lang="en">Cytoskeletal organization of human mesenchymal stem cells (MSC) changes during their osteogenic differentiation</title><author><name sortKey="Pablo Rodriguez, J" sort="Pablo Rodriguez, J" uniqKey="Pablo Rodriguez J" first="J." last="Pablo Rodríguez">J. Pablo Rodríguez</name><affiliation><mods:affiliation>Laboratorio de Biología Celular, INTA, Universidad de Chile, Casilla 138‐11, Santiago, Chile</mods:affiliation></affiliation><affiliation><mods:affiliation>E-mail: jprodrig@inta.cl</mods:affiliation></affiliation><affiliation><mods:affiliation>Correspondence address: Laboratorio de Biología Celular, INTA, Universidad de Chile, Casilla 138‐11, Santiago, Chile.</mods:affiliation></affiliation></author><author><name sortKey="Gonzalez, Mauricio" sort="Gonzalez, Mauricio" uniqKey="Gonzalez M" first="Mauricio" last="González">Mauricio González</name><affiliation><mods:affiliation>Laboratorio de Bioinformática y Expresión Génica, INTA, Universidad de Chile, Casilla 138‐11, Santiago, Chile</mods:affiliation></affiliation></author><author><name sortKey="Rios, Susana" sort="Rios, Susana" uniqKey="Rios S" first="Susana" last="Ríos">Susana Ríos</name><affiliation><mods:affiliation>Laboratorio de Biología Celular, INTA, Universidad de Chile, 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when="2004-11-01">2004-11-01</date></imprint><idno type="ISSN">0730-2312</idno></series></biblStruct></sourceDesc><seriesStmt><idno type="ISSN">0730-2312</idno></seriesStmt></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Actin</term><term>Actin cytoskeleton</term><term>Actin organization</term><term>Alexa fluor</term><term>Alkaline</term><term>Alkaline phosphatase activity</term><term>Basal</term><term>Basal conditions</term><term>Biochem</term><term>Biochemical markers</term><term>Bone marrow</term><term>Calcium deposition</term><term>Calcium phosphate deposition</term><term>Cell biochem</term><term>Cell biochem koukouritaki</term><term>Cell layer</term><term>Cell morphology</term><term>Cell shape</term><term>Characteristic phenotype</term><term>Chile</term><term>Cortical organization</term><term>Culture medium</term><term>Cytochalasin</term><term>Cytoskeletal</term><term>Cytoskeletal integrity</term><term>Cytoskeletal organization</term><term>Cytoskeleton</term><term>Cytoskeleton organization</term><term>Different donors</term><term>Differentiation</term><term>Differentiation markers</term><term>Entire cytoplasm</term><term>Human mesenchymal</term><term>Human mscs</term><term>Koukouritaki</term><term>Mesenchymal</term><term>Microtubule</term><term>Microtubule network</term><term>Morphological changes</term><term>Morphology</term><term>Msc</term><term>Nocodazole</term><term>Normal bone</term><term>Osteoblast</term><term>Osteoblastic</term><term>Osteogenic</term><term>Osteogenic conditions</term><term>Osteogenic differentiation</term><term>Osteogenic medium</term><term>Osteoporotic</term><term>Phenotype</term><term>Phosphatase</term><term>Rapid changes</term><term>Results show</term><term>Tubulin</term><term>Untreated</term><term>Untreated cells</term><term>Upper panels</term></keywords><keywords scheme="Teeft" xml:lang="en"><term>Actin</term><term>Actin cytoskeleton</term><term>Actin organization</term><term>Alexa fluor</term><term>Alkaline</term><term>Alkaline phosphatase activity</term><term>Basal</term><term>Basal conditions</term><term>Biochem</term><term>Biochemical markers</term><term>Bone marrow</term><term>Calcium deposition</term><term>Calcium phosphate deposition</term><term>Cell biochem</term><term>Cell biochem koukouritaki</term><term>Cell layer</term><term>Cell morphology</term><term>Cell shape</term><term>Characteristic phenotype</term><term>Chile</term><term>Cortical organization</term><term>Culture medium</term><term>Cytochalasin</term><term>Cytoskeletal</term><term>Cytoskeletal integrity</term><term>Cytoskeletal organization</term><term>Cytoskeleton</term><term>Cytoskeleton organization</term><term>Different donors</term><term>Differentiation</term><term>Differentiation markers</term><term>Entire cytoplasm</term><term>Human mesenchymal</term><term>Human mscs</term><term>Koukouritaki</term><term>Mesenchymal</term><term>Microtubule</term><term>Microtubule network</term><term>Morphological changes</term><term>Morphology</term><term>Msc</term><term>Nocodazole</term><term>Normal bone</term><term>Osteoblast</term><term>Osteoblastic</term><term>Osteogenic</term><term>Osteogenic conditions</term><term>Osteogenic differentiation</term><term>Osteogenic medium</term><term>Osteoporotic</term><term>Phenotype</term><term>Phosphatase</term><term>Rapid changes</term><term>Results show</term><term>Tubulin</term><term>Untreated</term><term>Untreated cells</term><term>Upper panels</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Human MSCs have been studied to define the mechanisms involved in normal bone remodeling and the regulation of osteogenesis. During osteogenic differentiation, MSCs change from their characteristic fibroblast‐like phenotype to near spherical shape. In this study, we analyzed the correlation between the organization of cytoskeleton of MSCs, changes in cell morphology, and the expression of specific markers (alkaline phosphatase activity and calcium deposition) of osteogenic differentiation. For osteoblastic differentiation, cells were cultured in a culture medium supplemented with 100 nM dexamethasone, 10 mM β‐ glycerophosphate, and 50 μg/ml ascorbic acid. The organization of microfilaments and microtubules was examined by inmunofluorescence using Alexa fluor 594 phalloidin and anti α‐tubulin monoclonal antibody. Cytochalasin D and nocodazole were used to alter reversibly the cytoskeleton dynamic. A remarkable change in cytoskeleton organization was observed in human MSCs during osteogenic differentiation. Actin cytoskeleton changed from a large number of thin, parallel microfilament bundles extending across the entire cytoplasm in undifferentiated MSCs to a few thick actin filament bundles located at the outermost periphery in differentiated cells. Under osteogenic culture conditions, a reversible reorganization of microfilaments induced by an initial treatment with cytochalasin D but not with nocodazole reduced the expression of differentiation markers, without affecting the final morphology of the cells. The results indicate that changes in the assembly and disassembly kinetics of microfilaments dynamic of actin network formation may be critical in supporting the osteogenic differentiation of human MSCs; also indicated that the organization of microtubules appears to have a regulatory role on the kinetic of this process. © 2004 Wiley‐Liss, Inc.</div></front></TEI><istex><corpusName>wiley</corpusName><keywords><teeft><json:string>osteogenic</json:string><json:string>msc</json:string><json:string>actin</json:string><json:string>cytoskeleton</json:string><json:string>cytochalasin</json:string><json:string>nocodazole</json:string><json:string>osteogenic differentiation</json:string><json:string>microtubule</json:string><json:string>cytoskeletal</json:string><json:string>phosphatase</json:string><json:string>osteoblast</json:string><json:string>basal</json:string><json:string>alkaline phosphatase activity</json:string><json:string>mesenchymal</json:string><json:string>osteogenic conditions</json:string><json:string>calcium deposition</json:string><json:string>biochem</json:string><json:string>cytoskeletal organization</json:string><json:string>cell shape</json:string><json:string>actin cytoskeleton</json:string><json:string>untreated cells</json:string><json:string>tubulin</json:string><json:string>phenotype</json:string><json:string>human mesenchymal</json:string><json:string>osteoporotic</json:string><json:string>koukouritaki</json:string><json:string>culture medium</json:string><json:string>osteoblastic</json:string><json:string>osteogenic medium</json:string><json:string>human mscs</json:string><json:string>cell morphology</json:string><json:string>untreated</json:string><json:string>biochemical markers</json:string><json:string>alkaline</json:string><json:string>cell layer</json:string><json:string>actin organization</json:string><json:string>microtubule network</json:string><json:string>cytoskeleton organization</json:string><json:string>differentiation</json:string><json:string>chile</json:string><json:string>morphology</json:string><json:string>alexa fluor</json:string><json:string>characteristic phenotype</json:string><json:string>cytoskeletal integrity</json:string><json:string>normal bone</json:string><json:string>upper panels</json:string><json:string>different donors</json:string><json:string>entire cytoplasm</json:string><json:string>basal conditions</json:string><json:string>calcium phosphate deposition</json:string><json:string>morphological changes</json:string><json:string>cortical organization</json:string><json:string>results show</json:string><json:string>rapid changes</json:string><json:string>cell biochem</json:string><json:string>bone marrow</json:string><json:string>cell biochem koukouritaki</json:string><json:string>differentiation markers</json:string></teeft></keywords><author><json:item><name>J. Pablo Rodríguez</name><affiliations><json:string>Laboratorio de Biología Celular, INTA, Universidad de Chile, Casilla 138‐11, Santiago, Chile</json:string><json:string>E-mail: jprodrig@inta.cl</json:string><json:string>Correspondence address: Laboratorio de Biología Celular, INTA, Universidad de Chile, Casilla 138‐11, Santiago, Chile.</json:string></affiliations></json:item><json:item><name>Mauricio González</name><affiliations><json:string>Laboratorio de Bioinformática y Expresión Génica, INTA, Universidad de Chile, Casilla 138‐11, Santiago, Chile</json:string></affiliations></json:item><json:item><name>Susana Ríos</name><affiliations><json:string>Laboratorio de Biología Celular, INTA, Universidad de Chile, Casilla 138‐11, Santiago, Chile</json:string></affiliations></json:item><json:item><name>Verónica Cambiazo</name><affiliations><json:string>Laboratorio de Bioinformática y Expresión Génica, INTA, Universidad de Chile, Casilla 138‐11, Santiago, Chile</json:string></affiliations></json:item></author><subject><json:item><lang><json:string>eng</json:string></lang><value>cytoskeleton</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>mesenchymal stem cells</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>osteogenic differentiation</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>actin</value></json:item><json:item><lang><json:string>eng</json:string></lang><value>tubulin</value></json:item></subject><articleId><json:string>JCB20234</json:string></articleId><arkIstex>ark:/67375/WNG-BXFKK9R7-7</arkIstex><language><json:string>eng</json:string></language><originalGenre><json:string>article</json:string></originalGenre><abstract>Human MSCs have been studied to define the mechanisms involved in normal bone remodeling and the regulation of osteogenesis. During osteogenic differentiation, MSCs change from their characteristic fibroblast‐like phenotype to near spherical shape. In this study, we analyzed the correlation between the organization of cytoskeleton of MSCs, changes in cell morphology, and the expression of specific markers (alkaline phosphatase activity and calcium deposition) of osteogenic differentiation. For osteoblastic differentiation, cells were cultured in a culture medium supplemented with 100 nM dexamethasone, 10 mM β‐ glycerophosphate, and 50 μg/ml ascorbic acid. The organization of microfilaments and microtubules was examined by inmunofluorescence using Alexa fluor 594 phalloidin and anti α‐tubulin monoclonal antibody. Cytochalasin D and nocodazole were used to alter reversibly the cytoskeleton dynamic. A remarkable change in cytoskeleton organization was observed in human MSCs during osteogenic differentiation. Actin cytoskeleton changed from a large number of thin, parallel microfilament bundles extending across the entire cytoplasm in undifferentiated MSCs to a few thick actin filament bundles located at the outermost periphery in differentiated cells. Under osteogenic culture conditions, a reversible reorganization of microfilaments induced by an initial treatment with cytochalasin D but not with nocodazole reduced the expression of differentiation markers, without affecting the final morphology of the cells. The results indicate that changes in the assembly and disassembly kinetics of microfilaments dynamic of actin network formation may be critical in supporting the osteogenic differentiation of human MSCs; also indicated that the organization of microtubules appears to have a regulatory role on the kinetic of this process. © 2004 Wiley‐Liss, Inc.</abstract><qualityIndicators><score>9.819</score><pdfWordCount>4819</pdfWordCount><pdfCharCount>32276</pdfCharCount><pdfVersion>1.3</pdfVersion><pdfPageCount>11</pdfPageCount><pdfPageSize>592 x 789 pts</pdfPageSize><refBibsNative>true</refBibsNative><abstractWordCount>258</abstractWordCount><abstractCharCount>1878</abstractCharCount><keywordCount>5</keywordCount></qualityIndicators><title>Cytoskeletal organization of human mesenchymal stem cells (MSC) changes during their osteogenic differentiation</title><pmid><json:string>15660416</json:string></pmid><genre><json:string>article</json:string></genre><host><title>Journal of Cellular Biochemistry</title><language><json:string>unknown</json:string></language><doi><json:string>10.1002/(ISSN)1097-4644</json:string></doi><issn><json:string>0730-2312</json:string></issn><eissn><json:string>1097-4644</json:string></eissn><publisherId><json:string>JCB</json:string></publisherId><volume>93</volume><issue>4</issue><pages><first>721</first><last>731</last><total>11</total></pages><genre><json:string>journal</json:string></genre><subject><json:item><value>Article</value></json:item></subject></host><namedEntities><unitex><date><json:string>2004</json:string></date><geogName></geogName><orgName><json:string>Hospital Sotero</json:string><json:string>Chile, Casilla</json:string><json:string>General Electric Medical Systems, Madison</json:string><json:string>MSC</json:string><json:string>Biochem</json:string><json:string>Hospital and INTA</json:string><json:string>Wiley-Liss, Inc</json:string><json:string>Cytoskeletal Organization</json:string><json:string>Molecular Probes Inc.</json:string></orgName><orgName_funder></orgName_funder><orgName_provider></orgName_provider><persName><json:string>O. Brunser</json:string><json:string>J. Cell</json:string><json:string>Alexa Fluor</json:string><json:string>In</json:string><json:string>J. Pablo</json:string><json:string>Mauricio Gonzalez</json:string><json:string>Microphotographs</json:string><json:string>Veronica Cambiazo</json:string><json:string>Insets</json:string></persName><placeName><json:string>Chile</json:string><json:string>Grunwald</json:string><json:string>Santiago</json:string><json:string>Japan</json:string><json:string>Tokyo</json:string></placeName><ref_url></ref_url><ref_bibl><json:string>Maccioni and Cambiazo, 1995</json:string><json:string>Perinpanayagam et al., 2001</json:string><json:string>Egan et al., 1991</json:string><json:string>Watanabe et al., 1997, 1998</json:string><json:string>Hughes-Fulford et al., 1992</json:string><json:string>Bellows et al., 1986</json:string><json:string>Nuttall et al., 1998</json:string><json:string>Koukouritaki et al., 1997</json:string><json:string>Gonzalez et al., 1998</json:string><json:string>Jaiswal et al., 1997</json:string><json:string>Takahashi et al., 1999</json:string><json:string>Hughes-Fulford and Lewis, 1996</json:string><json:string>Koukouritaki et al., 1996</json:string><json:string>Long et al., 1995</json:string><json:string>Yanaka et al., 2003</json:string><json:string>Haynesworth et al., 1992a,b</json:string><json:string>Lecanda et al., 1997</json:string><json:string>Spiegelman and Ginty, 1983</json:string><json:string>Luegmayr et al., 1996</json:string><json:string>Pavalko et al., 1998</json:string><json:string>Koukouritaki et al., 1999</json:string><json:string>Spiegelman and Farmer, 1982</json:string></ref_bibl><bibl></bibl></unitex></namedEntities><ark><json:string>ark:/67375/WNG-BXFKK9R7-7</json:string></ark><categories><wos><json:string>1 - 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vertebres: systeme osteoarticulaire, muscle strie</json:string></inist></categories><publicationDate>2004</publicationDate><copyrightDate>2004</copyrightDate><doi><json:string>10.1002/jcb.20234</json:string></doi><id>EEAEC74F03B67EADDC845D9F00ABA023094960BB</id><score>1</score><fulltext><json:item><extension>pdf</extension><original>true</original><mimetype>application/pdf</mimetype><uri>https://api.istex.fr/document/EEAEC74F03B67EADDC845D9F00ABA023094960BB/fulltext/pdf</uri></json:item><json:item><extension>zip</extension><original>false</original><mimetype>application/zip</mimetype><uri>https://api.istex.fr/document/EEAEC74F03B67EADDC845D9F00ABA023094960BB/fulltext/zip</uri></json:item><istex:fulltextTEI uri="https://api.istex.fr/document/EEAEC74F03B67EADDC845D9F00ABA023094960BB/fulltext/tei"><teiHeader><fileDesc><titleStmt><title level="a" type="main" xml:lang="en">Cytoskeletal organization of human mesenchymal stem cells (MSC) changes during their osteogenic differentiation</title></titleStmt><publicationStmt><authority>ISTEX</authority><publisher>Wiley Subscription Services, Inc., A Wiley Company</publisher><pubPlace>Hoboken</pubPlace><availability><licence>Copyright © 2004 Wiley‐Liss, Inc.</licence></availability><date type="published" when="2004-11-01"></date></publicationStmt><notesStmt><note type="content-type" subtype="article" source="article" scheme="https://content-type.data.istex.fr/ark:/67375/XTP-6N5SZHKN-D">article</note><note type="publication-type" subtype="journal" scheme="https://publication-type.data.istex.fr/ark:/67375/JMC-0GLKJH51-B">journal</note></notesStmt><sourceDesc><biblStruct type="article"><analytic><title level="a" type="main" xml:lang="en">Cytoskeletal organization of human mesenchymal stem cells (MSC) changes during their osteogenic differentiation</title><title level="a" type="short" xml:lang="en">Changes in Cytoskeleton in Human Mesenchymal Stem Cells</title><author xml:id="author-0000" role="corresp"><persName><forename type="first">J.</forename><surname>Pablo Rodríguez</surname></persName><email>jprodrig@inta.cl</email><affiliation>Laboratorio de Biología Celular, INTA, Universidad de Chile, Casilla 138‐11, Santiago, Chile<address><country key="CL"></country></address></affiliation><affiliation>Laboratorio de Biología Celular, INTA, Universidad de Chile, Casilla 138‐11, Santiago, Chile.</affiliation></author><author xml:id="author-0001"><persName><forename type="first">Mauricio</forename><surname>González</surname></persName><affiliation>Laboratorio de Bioinformática y Expresión Génica, INTA, Universidad de Chile, Casilla 138‐11, Santiago, Chile<address><country key="CL"></country></address></affiliation></author><author xml:id="author-0002"><persName><forename type="first">Susana</forename><surname>Ríos</surname></persName><affiliation>Laboratorio de Biología Celular, INTA, Universidad de Chile, Casilla 138‐11, Santiago, Chile<address><country key="CL"></country></address></affiliation></author><author xml:id="author-0003"><persName><forename type="first">Verónica</forename><surname>Cambiazo</surname></persName><affiliation>Laboratorio de Bioinformática y Expresión Génica, INTA, Universidad de Chile, Casilla 138‐11, Santiago, Chile<address><country key="CL"></country></address></affiliation></author><idno type="istex">EEAEC74F03B67EADDC845D9F00ABA023094960BB</idno><idno type="ark">ark:/67375/WNG-BXFKK9R7-7</idno><idno type="DOI">10.1002/jcb.20234</idno><idno type="unit">JCB20234</idno><idno type="toTypesetVersion">file:JCB.JCB20234.pdf</idno></analytic><monogr><title level="j" type="main">Journal of Cellular Biochemistry</title><title level="j" type="alt">JOURNAL OF CELLULAR BIOCHEMISTRY</title><idno type="pISSN">0730-2312</idno><idno type="eISSN">1097-4644</idno><idno type="book-DOI">10.1002/(ISSN)1097-4644</idno><idno type="book-part-DOI">10.1002/jcb.v93:4</idno><idno type="product">JCB</idno><imprint><biblScope unit="vol">93</biblScope><biblScope unit="issue">4</biblScope><biblScope unit="page" from="721">721</biblScope><biblScope unit="page" to="731">731</biblScope><biblScope unit="page-count">11</biblScope><publisher>Wiley Subscription Services, Inc., A Wiley Company</publisher><pubPlace>Hoboken</pubPlace><date type="published" when="2004-11-01"></date></imprint></monogr></biblStruct></sourceDesc></fileDesc><profileDesc><abstract xml:lang="en" style="main"><head>Abstract</head><p>Human MSCs have been studied to define the mechanisms involved in normal bone remodeling and the regulation of osteogenesis. During osteogenic differentiation, MSCs change from their characteristic fibroblast‐like phenotype to near spherical shape. In this study, we analyzed the correlation between the organization of cytoskeleton of MSCs, changes in cell morphology, and the expression of specific markers (alkaline phosphatase activity and calcium deposition) of osteogenic differentiation. For osteoblastic differentiation, cells were cultured in a culture medium supplemented with 100 nM dexamethasone, 10 mM β‐ glycerophosphate, and 50 μg/ml ascorbic acid. The organization of microfilaments and microtubules was examined by inmunofluorescence using Alexa fluor 594 phalloidin and anti α‐tubulin monoclonal antibody. Cytochalasin D and nocodazole were used to alter reversibly the cytoskeleton dynamic. A remarkable change in cytoskeleton organization was observed in human MSCs during osteogenic differentiation. Actin cytoskeleton changed from a large number of thin, parallel microfilament bundles extending across the entire cytoplasm in undifferentiated MSCs to a few thick actin filament bundles located at the outermost periphery in differentiated cells. Under osteogenic culture conditions, a reversible reorganization of microfilaments induced by an initial treatment with cytochalasin D but not with nocodazole reduced the expression of differentiation markers, without affecting the final morphology of the cells. The results indicate that changes in the assembly and disassembly kinetics of microfilaments dynamic of actin network formation may be critical in supporting the osteogenic differentiation of human MSCs; also indicated that the organization of microtubules appears to have a regulatory role on the kinetic of this process. © 2004 Wiley‐Liss, Inc.</p></abstract><textClass><keywords xml:lang="en"><term xml:id="kwd1">cytoskeleton</term><term xml:id="kwd2">mesenchymal stem cells</term><term xml:id="kwd3">osteogenic differentiation</term><term xml:id="kwd4">actin</term><term xml:id="kwd5">tubulin</term></keywords><keywords rend="articleCategory"><term>Article</term></keywords><keywords rend="tocHeading1"><term>Articles</term></keywords></textClass><langUsage><language ident="en"></language></langUsage></profileDesc></teiHeader></istex:fulltextTEI><json:item><extension>txt</extension><original>false</original><mimetype>text/plain</mimetype><uri>https://api.istex.fr/document/EEAEC74F03B67EADDC845D9F00ABA023094960BB/fulltext/txt</uri></json:item></fulltext><metadata><istex:metadataXml wicri:clean="Wiley, elements deleted: body"><istex:xmlDeclaration>version="1.0" encoding="UTF-8" standalone="yes"</istex:xmlDeclaration><istex:document><component version="2.0" type="serialArticle" xml:lang="en"><header><publicationMeta level="product"><publisherInfo><publisherName>Wiley Subscription Services, Inc., A Wiley Company</publisherName><publisherLoc>Hoboken</publisherLoc></publisherInfo><doi registered="yes">10.1002/(ISSN)1097-4644</doi><issn type="print">0730-2312</issn><issn type="electronic">1097-4644</issn><idGroup><id type="product" value="JCB"></id></idGroup><titleGroup><title type="main" xml:lang="en" sort="JOURNAL OF CELLULAR BIOCHEMISTRY">Journal of Cellular Biochemistry</title><title type="short">J. 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Actin cytoskeleton changed from a large number of thin, parallel microfilament bundles extending across the entire cytoplasm in undifferentiated MSCs to a few thick actin filament bundles located at the outermost periphery in differentiated cells. Under osteogenic culture conditions, a reversible reorganization of microfilaments induced by an initial treatment with cytochalasin D but not with nocodazole reduced the expression of differentiation markers, without affecting the final morphology of the cells. The results indicate that changes in the assembly and disassembly kinetics of microfilaments dynamic of actin network formation may be critical in supporting the osteogenic differentiation of human MSCs; also indicated that the organization of microtubules appears to have a regulatory role on the kinetic of this process. © 2004 Wiley‐Liss, Inc.</p></abstract></abstractGroup></contentMeta></header></component></istex:document></istex:metadataXml><mods version="3.6"><titleInfo lang="en"><title>Cytoskeletal organization of human mesenchymal stem cells (MSC) changes during their osteogenic differentiation</title></titleInfo><titleInfo type="abbreviated" lang="en"><title>Changes in Cytoskeleton in Human Mesenchymal Stem Cells</title></titleInfo><titleInfo type="alternative" contentType="CDATA" lang="en"><title>Cytoskeletal organization of human mesenchymal stem cells (MSC) changes during their osteogenic differentiation</title></titleInfo><name type="personal"><namePart type="given">J.</namePart><namePart type="family">Pablo Rodríguez</namePart><affiliation>Laboratorio de Biología Celular, INTA, Universidad de Chile, Casilla 138‐11, Santiago, Chile</affiliation><affiliation>E-mail: jprodrig@inta.cl</affiliation><affiliation>Correspondence address: Laboratorio de Biología Celular, INTA, Universidad de Chile, Casilla 138‐11, Santiago, Chile.</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Mauricio</namePart><namePart type="family">González</namePart><affiliation>Laboratorio de Bioinformática y Expresión Génica, INTA, Universidad de Chile, Casilla 138‐11, Santiago, Chile</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Susana</namePart><namePart type="family">Ríos</namePart><affiliation>Laboratorio de Biología Celular, INTA, Universidad de Chile, Casilla 138‐11, Santiago, Chile</affiliation><role><roleTerm type="text">author</roleTerm></role></name><name type="personal"><namePart type="given">Verónica</namePart><namePart type="family">Cambiazo</namePart><affiliation>Laboratorio de Bioinformática y Expresión Génica, INTA, Universidad de Chile, Casilla 138‐11, Santiago, Chile</affiliation><role><roleTerm type="text">author</roleTerm></role></name><typeOfResource>text</typeOfResource><genre type="article" displayLabel="article" authority="ISTEX" authorityURI="https://content-type.data.istex.fr" valueURI="https://content-type.data.istex.fr/ark:/67375/XTP-6N5SZHKN-D">article</genre><originInfo><publisher>Wiley Subscription Services, Inc., A Wiley Company</publisher><place><placeTerm type="text">Hoboken</placeTerm></place><dateIssued encoding="w3cdtf">2004-11-01</dateIssued><dateCaptured encoding="w3cdtf">2004-04-01</dateCaptured><dateValid encoding="w3cdtf">2004-06-11</dateValid><copyrightDate encoding="w3cdtf">2004</copyrightDate></originInfo><language><languageTerm type="code" authority="rfc3066">en</languageTerm><languageTerm type="code" authority="iso639-2b">eng</languageTerm></language><physicalDescription><extent unit="figures">3</extent><extent unit="tables">0</extent><extent unit="references">31</extent><extent unit="words">5322</extent></physicalDescription><abstract lang="en">Human MSCs have been studied to define the mechanisms involved in normal bone remodeling and the regulation of osteogenesis. During osteogenic differentiation, MSCs change from their characteristic fibroblast‐like phenotype to near spherical shape. In this study, we analyzed the correlation between the organization of cytoskeleton of MSCs, changes in cell morphology, and the expression of specific markers (alkaline phosphatase activity and calcium deposition) of osteogenic differentiation. For osteoblastic differentiation, cells were cultured in a culture medium supplemented with 100 nM dexamethasone, 10 mM β‐ glycerophosphate, and 50 μg/ml ascorbic acid. The organization of microfilaments and microtubules was examined by inmunofluorescence using Alexa fluor 594 phalloidin and anti α‐tubulin monoclonal antibody. Cytochalasin D and nocodazole were used to alter reversibly the cytoskeleton dynamic. A remarkable change in cytoskeleton organization was observed in human MSCs during osteogenic differentiation. Actin cytoskeleton changed from a large number of thin, parallel microfilament bundles extending across the entire cytoplasm in undifferentiated MSCs to a few thick actin filament bundles located at the outermost periphery in differentiated cells. Under osteogenic culture conditions, a reversible reorganization of microfilaments induced by an initial treatment with cytochalasin D but not with nocodazole reduced the expression of differentiation markers, without affecting the final morphology of the cells. 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