title: An in vitro model of skeletal muscle volume regulation.
abstract: BACKGROUND
  Hypertonic media causes cells to shrink due to water loss through aquaporin channels. After acute shrinkage, cells either regulate their volume or, alternatively, undergo a number of metabolic changes which ultimately lead to cell death. In many cell types, hypertonic shrinkage is followed by apoptosis. Due to the complex 3D morphology of skeletal muscle and the difficulty in obtaining isolated human tissue, we have begun skeletal muscle volume regulation studies using the human skeletal muscle cell line TE671RD. In this study we investigated whether hypertonic challenge of the human skeletal muscle cell line TE671RD triggered cell death or evoked a cell volume recovery response.
  
  
  METHODS
  The cellular volume of TE671RD cells was calculated from the 2D surface area. Cell death was assessed by both the trypan blue live/dead assay and the TUNEL assay.
  
  
  RESULTS
  Medium osmolality was increased by addition of up to 200 mM sucrose. Addition of 200 mM sucrose resulted in mean cell shrinkage of 44±1% after 30 mins. At later time points (2 and 4 hrs) two separate cell subpopulations with differing mean cell volume became apparent. The first subpopulation (15±2% of the total cell number) continued to shrink whereas the second subpopulation had an increased cell volume. Cell death was observed in a small proportion of cells (approximately 6-8%).
  
  
  CONCLUSIONS
  We have established that a substantial proportion of TE671RD cells respond to hypertonic challenge with RVI, but that these cells are resistant to hypertonicity triggered cell death.
authors.affiliation: Department of Musculoskeletal Biology, Institute of Ageing and Chronic Disease, University of Liverpool, Liverpool, United Kingdom.
authors.forename: Anna
authors.initials: A
authors.lastname: Wibberley
authors.affiliation: Department of Musculoskeletal Biology, Institute of Ageing and Chronic Disease, University of Liverpool, Liverpool, United Kingdom.
authors.forename: Caroline A
authors.initials: CA
authors.lastname: Staunton
authors.affiliation: Department of Musculoskeletal Biology, Institute of Ageing and Chronic Disease, University of Liverpool, Liverpool, United Kingdom.
authors.forename: Claire H
authors.initials: CH
authors.lastname: Feetham
authors.affiliation: Laboratory of Cell Physiology, Institute of Cytology, Russian Academy of Sciences, St-Petersburg, Russia.
authors.forename: Alexey A
authors.initials: AA
authors.lastname: Vereninov
authors.affiliation: Department of Musculoskeletal Biology, Institute of Ageing and Chronic Disease, University of Liverpool, Liverpool, United Kingdom.
authors.forename: Richard
authors.initials: R
authors.lastname: Barrett-Jolley
mesh_terms: D017209:Apoptosis / Q000187:drug effects; D002460:Cell Line; D048429:Cell Size / Q000187:drug effects; D006801:Humans; D020287:In Situ Nick-End Labeling; D018482:Muscle, Skeletal / Q000166:cytology*; D009994:Osmolar Concentration; D013395:Sucrose / Q000494:pharmacology
chemical_list: D013395:Sucrose