Serum from differently exercised subjects induces myogenic differentiation in LHCN-M2 human myoblasts

J Sports Sci. 2018 Jul;36(14):1630-1639. doi: 10.1080/02640414.2017.1407232. Epub 2017 Nov 21.

Abstract

Myogenesis is the formation of muscle tissue from muscle precursor cells. Physical exercise induces satellite cell activation in muscle. Currently, C2C12 murine myoblast cells are used to study myogenic differentiation. Herein, we evaluated whether human LHCN-M2 myoblasts can differentiate into mature myotubes and express early (myotube formation, creatine kinase activity and myogenin) and late (MyHC-β) muscle-specific markers when cultured in differentiation medium (DM) for 2, 4 and 7 days. We demonstrate that treatment of LHCN-M2 cells with DM supplemented with 0.5% serum from long-term (3 years) differently exercised subjects for 4 days induced myotube formation and significantly increased the early (creatine kinase activity and myogenin) and late (MyHC-β expression) differentiation markers versus cells treated with serum from untrained subjects. Interestingly, serum from aerobic exercised subjects (swimming) had a greater positive effect on late-differentiation marker (MyHC-β) expression than serum from anaerobic (body building) or from mixed exercised (soccer and volleyball) subjects. Moreover, p62and anti-apoptotic Bcl-2 protein expression was lower in LHCN-M2 cells cultured with human sera from differently exercised subjectst han in cells cultured with DM. In conclusion, LHCN-M2 human myoblasts represent a species-specific system with which to study human myogenic differentiation induced by serum from differently exercised subjects.

Keywords: Exercise; LHCN-M2 cells; human myoblasts; human serum; myogenic differentiation; myosin heavy chain-β (MyHC-β).

MeSH terms

  • Adult
  • Apoptosis / physiology
  • Autophagy / physiology
  • Cardiac Myosins / genetics
  • Cardiac Myosins / metabolism
  • Cell Culture Techniques*
  • Cell Differentiation / physiology*
  • Cell Line
  • Creatine Kinase / metabolism
  • Culture Media
  • Exercise / physiology*
  • Gene Expression
  • Humans
  • Muscle Development / physiology*
  • Muscle Fibers, Skeletal / physiology
  • Myoblasts / physiology*
  • Myogenin / metabolism
  • Myosin Heavy Chains / genetics
  • Myosin Heavy Chains / metabolism
  • RNA, Messenger / genetics
  • Serum
  • Young Adult

Substances

  • Culture Media
  • MYH7 protein, human
  • Myogenin
  • RNA, Messenger
  • Creatine Kinase
  • Cardiac Myosins
  • Myosin Heavy Chains