title: Skeletal muscle characteristics are preserved in hTERT/cdk4 human myogenic cell lines.
abstract: hTERT/cdk4 immortalized myogenic human cell lines represent an important tool for skeletal muscle research, being used as therapeutically pertinent models of various neuromuscular disorders and in numerous fundamental studies of muscle cell function. However, the cell cycle is linked to other cellular processes such as integrin regulation, the PI3K/Akt pathway, and microtubule stability, raising the question as to whether genetic modification related to the cell cycle results in secondary effects that could undermine the validity of these cell models.
  
  
  
  Here we subjected five healthy and disease muscle cell isolates to transcriptomic analysis, comparing immortalized lines with their parent primary populations in both differentiated and undifferentiated states, and testing their myogenic character by comparison with non-myogenic (CD56-negative) cells. Principal component analysis of global gene expression showed tight clustering of immortalized myoblasts to their parent primary populations, with clean separation from the non-myogenic reference. Comparison was made to publicly available transcriptomic data from studies of muscle human pathology, cell, and animal models, including to derive a consensus set of genes previously shown to have altered regulation during myoblast differentiation. Hierarchical clustering of samples based on gene expression of this consensus set showed that immortalized lines retained the myogenic expression patterns of their parent primary populations. Of 2784 canonical pathways and gene ontology terms tested by gene set enrichment analysis, none were significantly enriched in immortalized compared to primary cell populations. We observed, at the whole transcriptome level, a strong signature of cell cycle shutdown associated with senescence in one primary myoblast population, whereas its immortalized clone was protected.
  
  
  
  Immortalization had no observed effect on the myogenic cascade or on any other cellular processes, and it was protective against the systems level effects of senescence that are observed at higher division counts of primary cells.
authors.affiliation: INSERM, CNRS, Institute of Myology, Center of Research in Myology, Sorbonne Universities, UPMC Univ Paris 6, Paris, France.
authors.forename: Matthew
authors.initials: M
authors.lastname: Thorley
authors.affiliation: INSERM, CNRS, Institute of Myology, Center of Research in Myology, Sorbonne Universities, UPMC Univ Paris 6, Paris, France.|Northern Ireland Centre for Stratified Medicine, Altnagelvin Hospital Campus, Ulster University, Londonderry, Northern Ireland UK.
authors.forename: Stéphanie
authors.initials: S
authors.lastname: Duguez
authors.affiliation: Department of Life Sciences, Center for Genome Research, University of Modena and Reggio Emilia, Modena, Italy.
authors.forename: Emilia Maria Cristina
authors.initials: EMC
authors.lastname: Mazza
authors.affiliation: Department of Life Sciences, Center for Genome Research, University of Modena and Reggio Emilia, Modena, Italy.
authors.forename: Sara
authors.initials: S
authors.lastname: Valsoni
authors.affiliation: INSERM, CNRS, Institute of Myology, Center of Research in Myology, Sorbonne Universities, UPMC Univ Paris 6, Paris, France.
authors.forename: Anne
authors.initials: A
authors.lastname: Bigot
authors.affiliation: INSERM, CNRS, Institute of Myology, Center of Research in Myology, Sorbonne Universities, UPMC Univ Paris 6, Paris, France.
authors.forename: Kamel
authors.initials: K
authors.lastname: Mamchaoui
authors.affiliation: Research Center for Genetic Medicine, Children's National Medical Center, Washington, DC 20010 USA.
authors.forename: Brennan
authors.initials: B
authors.lastname: Harmon
authors.affiliation: NIHR Biomedical Research Centre, Institute of Child Health, University College London, 30 Guilford Street, London, UK.
authors.forename: Thomas
authors.initials: T
authors.lastname: Voit
authors.affiliation: INSERM, CNRS, Institute of Myology, Center of Research in Myology, Sorbonne Universities, UPMC Univ Paris 6, Paris, France.
authors.forename: Vincent
authors.initials: V
authors.lastname: Mouly
authors.affiliation: INSERM, CNRS, Institute of Myology, Center of Research in Myology, Sorbonne Universities, UPMC Univ Paris 6, Paris, France.|Northern Ireland Centre for Stratified Medicine, Altnagelvin Hospital Campus, Ulster University, Londonderry, Northern Ireland UK.
authors.forename: William
authors.identifier: 0000-0003-2239-9094
authors.initials: W
authors.lastname: Duddy
mesh_terms: D002460:Cell Line; D002478:Cells, Cultured; D016922:Cellular Senescence; D051358:Cyclin-Dependent Kinase 4 / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D024510:Muscle Development; D032446:Myoblasts / Q000166:cytology / Q000378:metabolism*; D019098:Telomerase / Q000235:genetics* / Q000378:metabolism; D059467:Transcriptome
chemical_list: C495900:CDK4 protein, human; D051358:Cyclin-Dependent Kinase 4; C509186:TERT protein, human; D019098:Telomerase