A cellular model system of differentiated human myotubes
The aim of this study was to select an effective and stable protocol for the differentiation of human satellite cells (Sc) and to identify the optimal time period for the experimental use of differentiated human Sc‐cultures. In order to identify the differentiation conditions which give a good survi...
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Veröffentlicht in: | APMIS : acta pathologica, microbiologica et immunologica Scandinavica microbiologica et immunologica Scandinavica, 2001-11, Vol.109 (11), p.735-744 |
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creator | GASTER, M. KRISTENSEN, S. R. BECK-NIELSEN, H. SCHRØDER, H. D. |
description | The aim of this study was to select an effective and stable protocol for the differentiation of human satellite cells (Sc) and to identify the optimal time period for the experimental use of differentiated human Sc‐cultures. In order to identify the differentiation conditions which give a good survival of myotubes and a high grade of differentiation, Sc‐cultures were induced to differentiate in media supplemented with either 2% fetal calf serum (FCS) 2% horse serum (HS) or 10% HS. Based on higher CK‐activities in cultures differentiating in FCS‐supplemented media compared to horse sera, fetal calf serum was chosen to induce differentiation. The ATP, DNA and protein content increased during the first 4 days after induction of differentiation and was followed by a period with minor changes. The maximal differences of ATP, DNA and protein between days 4–10 were evaluated and the differences in the three components were found to be less than 20% of the average value with a certainity of more than 0.9. Day 8‐myotubes were investigated morphologically and were found immunoreactive for fast myosin, and expressed areas with clear cross striation. We recommend the use of differentiated Sc‐cultures in the period from day 4 to 8 after induction of differentiation as only minor differentation‐related changes will take place in the cells during this period of time. |
doi_str_mv | 10.1034/j.1600-0463.2001.d01-140.x |
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R. ; BECK-NIELSEN, H. ; SCHRØDER, H. D.</creator><creatorcontrib>GASTER, M. ; KRISTENSEN, S. R. ; BECK-NIELSEN, H. ; SCHRØDER, H. D.</creatorcontrib><description>The aim of this study was to select an effective and stable protocol for the differentiation of human satellite cells (Sc) and to identify the optimal time period for the experimental use of differentiated human Sc‐cultures. In order to identify the differentiation conditions which give a good survival of myotubes and a high grade of differentiation, Sc‐cultures were induced to differentiate in media supplemented with either 2% fetal calf serum (FCS) 2% horse serum (HS) or 10% HS. Based on higher CK‐activities in cultures differentiating in FCS‐supplemented media compared to horse sera, fetal calf serum was chosen to induce differentiation. The ATP, DNA and protein content increased during the first 4 days after induction of differentiation and was followed by a period with minor changes. The maximal differences of ATP, DNA and protein between days 4–10 were evaluated and the differences in the three components were found to be less than 20% of the average value with a certainity of more than 0.9. Day 8‐myotubes were investigated morphologically and were found immunoreactive for fast myosin, and expressed areas with clear cross striation. We recommend the use of differentiated Sc‐cultures in the period from day 4 to 8 after induction of differentiation as only minor differentation‐related changes will take place in the cells during this period of time.</description><identifier>ISSN: 0903-4641</identifier><identifier>EISSN: 1600-0463</identifier><identifier>DOI: 10.1034/j.1600-0463.2001.d01-140.x</identifier><identifier>PMID: 11900052</identifier><language>eng</language><publisher>Copenhagen: Munksgaard International Publishers</publisher><subject>Adenosine Triphosphate - metabolism ; Bacterial diseases ; Biological and medical sciences ; Cell culture ; Cell Differentiation ; Cells, Cultured ; Creatine Kinase - metabolism ; Creatine Kinase, MM Form ; differentiation ; DNA - metabolism ; Experimental bacterial diseases and models ; Human viral diseases ; Humans ; Immunohistochemistry ; Infectious diseases ; Isoenzymes - metabolism ; Medical sciences ; Microscopy, Electron ; Models, Biological ; Muscle Proteins - metabolism ; Muscle, Skeletal - cytology ; Muscle, Skeletal - metabolism ; Myosins - metabolism ; satellite cells ; skeletal muscle ; Stem Cells - cytology ; Stem Cells - metabolism ; Viral diseases ; Viral diseases of the lymphoid tissue and the blood. 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R.</creatorcontrib><creatorcontrib>BECK-NIELSEN, H.</creatorcontrib><creatorcontrib>SCHRØDER, H. D.</creatorcontrib><title>A cellular model system of differentiated human myotubes</title><title>APMIS : acta pathologica, microbiologica et immunologica Scandinavica</title><addtitle>APMIS</addtitle><description>The aim of this study was to select an effective and stable protocol for the differentiation of human satellite cells (Sc) and to identify the optimal time period for the experimental use of differentiated human Sc‐cultures. In order to identify the differentiation conditions which give a good survival of myotubes and a high grade of differentiation, Sc‐cultures were induced to differentiate in media supplemented with either 2% fetal calf serum (FCS) 2% horse serum (HS) or 10% HS. Based on higher CK‐activities in cultures differentiating in FCS‐supplemented media compared to horse sera, fetal calf serum was chosen to induce differentiation. The ATP, DNA and protein content increased during the first 4 days after induction of differentiation and was followed by a period with minor changes. The maximal differences of ATP, DNA and protein between days 4–10 were evaluated and the differences in the three components were found to be less than 20% of the average value with a certainity of more than 0.9. Day 8‐myotubes were investigated morphologically and were found immunoreactive for fast myosin, and expressed areas with clear cross striation. We recommend the use of differentiated Sc‐cultures in the period from day 4 to 8 after induction of differentiation as only minor differentation‐related changes will take place in the cells during this period of time.</description><subject>Adenosine Triphosphate - metabolism</subject><subject>Bacterial diseases</subject><subject>Biological and medical sciences</subject><subject>Cell culture</subject><subject>Cell Differentiation</subject><subject>Cells, Cultured</subject><subject>Creatine Kinase - metabolism</subject><subject>Creatine Kinase, MM Form</subject><subject>differentiation</subject><subject>DNA - metabolism</subject><subject>Experimental bacterial diseases and models</subject><subject>Human viral diseases</subject><subject>Humans</subject><subject>Immunohistochemistry</subject><subject>Infectious diseases</subject><subject>Isoenzymes - metabolism</subject><subject>Medical sciences</subject><subject>Microscopy, Electron</subject><subject>Models, Biological</subject><subject>Muscle Proteins - metabolism</subject><subject>Muscle, Skeletal - cytology</subject><subject>Muscle, Skeletal - metabolism</subject><subject>Myosins - metabolism</subject><subject>satellite cells</subject><subject>skeletal muscle</subject><subject>Stem Cells - cytology</subject><subject>Stem Cells - metabolism</subject><subject>Viral diseases</subject><subject>Viral diseases of the lymphoid tissue and the blood. Aids</subject><issn>0903-4641</issn><issn>1600-0463</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2001</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqVkVtv1DAQRq2Kii6Fv4AiJHhLOo5vCW-rBbZAuUmtQLxY40tElmRT7ETd_fd1tKv2GcmSH3zm88wZQl5RKCgwfrEpqATIgUtWlAC0cEBzyqHYnZDFw9MTsoAaWM4lp2fkWYybhJaVVE_JGaU1AIhyQaplZn3XTR2GrB-c77K4j6Pvs6HJXNs0Pvjt2OLoXfZn6nGb9fthnIyPz8lpg130L473Obn58P56dZlffVt_XC2vcssVY3ltkNpaqlpahgIatKI00iAKWxthSu4o41QwJV3FHBPc-RopGmsrWimD7Jy8OeTehuHf5OOo-zbOLePWD1PUKs0kQEEC3x5AG4YYg2_0bWh7DHtNQc_e9EbPcvQsR8_edPKmkze9S8Uvj79MpvfusfQoKgGvjwBGi10TcGvb-MhxKlUaI3HvDtxd2_n9f7Sgl9-_pAM1TWCKyQ8xbVrG7iEGw18tFVNC__y61r9WP7j4tP6sf7N7Qx-ZvQ</recordid><startdate>200111</startdate><enddate>200111</enddate><creator>GASTER, M.</creator><creator>KRISTENSEN, S. R.</creator><creator>BECK-NIELSEN, H.</creator><creator>SCHRØDER, H. D.</creator><general>Munksgaard International Publishers</general><general>Blackwell</general><scope>BSCLL</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>200111</creationdate><title>A cellular model system of differentiated human myotubes</title><author>GASTER, M. ; KRISTENSEN, S. R. ; BECK-NIELSEN, H. ; SCHRØDER, H. D.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c4733-9ba1c96796c3a50fac52b6baa5c9b5b24d13415376d83d354de9a1abcc8187ba3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2001</creationdate><topic>Adenosine Triphosphate - metabolism</topic><topic>Bacterial diseases</topic><topic>Biological and medical sciences</topic><topic>Cell culture</topic><topic>Cell Differentiation</topic><topic>Cells, Cultured</topic><topic>Creatine Kinase - metabolism</topic><topic>Creatine Kinase, MM Form</topic><topic>differentiation</topic><topic>DNA - metabolism</topic><topic>Experimental bacterial diseases and models</topic><topic>Human viral diseases</topic><topic>Humans</topic><topic>Immunohistochemistry</topic><topic>Infectious diseases</topic><topic>Isoenzymes - metabolism</topic><topic>Medical sciences</topic><topic>Microscopy, Electron</topic><topic>Models, Biological</topic><topic>Muscle Proteins - metabolism</topic><topic>Muscle, Skeletal - cytology</topic><topic>Muscle, Skeletal - metabolism</topic><topic>Myosins - metabolism</topic><topic>satellite cells</topic><topic>skeletal muscle</topic><topic>Stem Cells - cytology</topic><topic>Stem Cells - metabolism</topic><topic>Viral diseases</topic><topic>Viral diseases of the lymphoid tissue and the blood. Aids</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>GASTER, M.</creatorcontrib><creatorcontrib>KRISTENSEN, S. R.</creatorcontrib><creatorcontrib>BECK-NIELSEN, H.</creatorcontrib><creatorcontrib>SCHRØDER, H. D.</creatorcontrib><collection>Istex</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>APMIS : acta pathologica, microbiologica et immunologica Scandinavica</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>GASTER, M.</au><au>KRISTENSEN, S. R.</au><au>BECK-NIELSEN, H.</au><au>SCHRØDER, H. D.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>A cellular model system of differentiated human myotubes</atitle><jtitle>APMIS : acta pathologica, microbiologica et immunologica Scandinavica</jtitle><addtitle>APMIS</addtitle><date>2001-11</date><risdate>2001</risdate><volume>109</volume><issue>11</issue><spage>735</spage><epage>744</epage><pages>735-744</pages><issn>0903-4641</issn><eissn>1600-0463</eissn><abstract>The aim of this study was to select an effective and stable protocol for the differentiation of human satellite cells (Sc) and to identify the optimal time period for the experimental use of differentiated human Sc‐cultures. In order to identify the differentiation conditions which give a good survival of myotubes and a high grade of differentiation, Sc‐cultures were induced to differentiate in media supplemented with either 2% fetal calf serum (FCS) 2% horse serum (HS) or 10% HS. Based on higher CK‐activities in cultures differentiating in FCS‐supplemented media compared to horse sera, fetal calf serum was chosen to induce differentiation. The ATP, DNA and protein content increased during the first 4 days after induction of differentiation and was followed by a period with minor changes. The maximal differences of ATP, DNA and protein between days 4–10 were evaluated and the differences in the three components were found to be less than 20% of the average value with a certainity of more than 0.9. Day 8‐myotubes were investigated morphologically and were found immunoreactive for fast myosin, and expressed areas with clear cross striation. We recommend the use of differentiated Sc‐cultures in the period from day 4 to 8 after induction of differentiation as only minor differentation‐related changes will take place in the cells during this period of time.</abstract><cop>Copenhagen</cop><pub>Munksgaard International Publishers</pub><pmid>11900052</pmid><doi>10.1034/j.1600-0463.2001.d01-140.x</doi><tpages>10</tpages></addata></record> |
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subjects | Adenosine Triphosphate - metabolism Bacterial diseases Biological and medical sciences Cell culture Cell Differentiation Cells, Cultured Creatine Kinase - metabolism Creatine Kinase, MM Form differentiation DNA - metabolism Experimental bacterial diseases and models Human viral diseases Humans Immunohistochemistry Infectious diseases Isoenzymes - metabolism Medical sciences Microscopy, Electron Models, Biological Muscle Proteins - metabolism Muscle, Skeletal - cytology Muscle, Skeletal - metabolism Myosins - metabolism satellite cells skeletal muscle Stem Cells - cytology Stem Cells - metabolism Viral diseases Viral diseases of the lymphoid tissue and the blood. Aids |
title | A cellular model system of differentiated human myotubes |
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