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
Hauptverfasser: GASTER, M., KRISTENSEN, S. R., BECK-NIELSEN, H., SCHRØDER, H. D.
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container_title APMIS : acta pathologica, microbiologica et immunologica Scandinavica
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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.
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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. 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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. 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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. 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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. 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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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