title: A semi-automated programme for tracking myoblast migration following mechanical damage: manipulation by chemical inhibitors.
abstract: BACKGROUND
  Potential roles for undifferentiated skeletal muscle stem cells or satellite cells in muscle hypertrophy and repair have been reported, however, the capacity, the mode and the mechanisms underpinning migration have not been investigated. We hypothesised that damaged skeletal myoblasts would elicit a mesenchymal-like migratory response, which could be precisely tracked and subsequently manipulated.
  
  
  METHODS
  We therefore established a model of mechanical damage and developed a MATLAB(TM) tool to measure the migratory capacity of myoblasts in a non-subjective manner.
  
  
  RESULTS
  Basal migration following damage was highly directional, with total migration distances of 948μm ± 239μm being recorded (average 0-24 hour distances: 491μm ± 113μm and 24-48 hour distances: 460μm ± 218μm). Pharmacological inhibition of MEK or PI3-K using PD98059 (20μM) or LY294002 (5μm), resulted in significant reduction of overall cell migration distances of 38% (p<0.001) and 39.5% (p<0.0004), respectively. Using the semi-automated cell tracking using MATLAB(TM) program we validated that not only was migration distance reduced as a consequence of reduced cell velocity, but critically also as a result of altered directionality of migration.
  
  
  CONCLUSIONS
  These studies demonstrate that murine myoblasts in culture migrate and provide a good model for studying responsiveness to damage in vitro. They illustrate for the first time the powerful tool that MATLAB(TM) provides in determining that both velocity and directional capacity influence the migratory potential of cellular movement with obvious implications for homing and for metastases.
authors.affiliation: School of Healthcare Science, Institute for Biomedical Research Into Human Movement and Health (IRM), MMU, Oxford Road, Manchester, UK. n.al-shanti@mmu.ac.uk
authors.forename: Nasser
authors.initials: N
authors.lastname: Al-Shanti
authors.forename: Steve H
authors.initials: SH
authors.lastname: Faulkner
authors.forename: Amarjit
authors.initials: A
authors.lastname: Saini
authors.forename: Ian
authors.initials: I
authors.lastname: Loram
authors.forename: Claire E
authors.initials: CE
authors.lastname: Stewart
mesh_terms: D000818:Animals; D001331:Automation; D002460:Cell Line; D002465:Cell Movement; D003470:Culture Media; D051379:Mice; D018482:Muscle, Skeletal / Q000166:cytology / Q000187:drug effects; D032446:Myoblasts / Q000166:cytology*
chemical_list: D003470:Culture Media