Investigating the effects of 5.5 mmoL vs 25 mmoL glucose concentration in culture media on LHCN-M2 cell viability, proliferation, metabolism and differentiation

Introduction: In vitro skeletal muscle cell models are vitally important for investigating the molecular mechanisms of skeletal muscle in metabolic and endocrine diseases, such as obesity and type 2 diabetes. Culture media for skeletal muscle cells can often contain glucose concentrations (GC) five times higher than what’s considered normal in fasting human plasma, thus is not representative of the in vivo environment. Hyperglycaemia in culture media may negatively impact metabolic function, by creating a model of cell toxicity that’s representative of diseases such as diabetes mellitus. The aim of these experiments was to determine the impact of media containing GC of 5.5 mmol (physiological) vs 25 mmol (supraphysiological) on cell viability, proliferation, ATP production and differentiation in human LHCN-m2 myoblasts.

Methods: LHCN-m2 myoblasts were cultured in 5.5 mmol or 25 mmol glucose growth media and cell viability, ATP production, and proliferation were determined. Differentiation of LHCN-m2 myoblasts into multinucleated myotubes was induced by reducing levels of human serum within the culture media and analysed by immunofluorescence following 10 days of differentiation.

Results: We observed no differences in the viability, proliferation or basal ATP production rates of LHCN-m2 cells grown in 5.5 mmol compared to 25 mmol glucose (P> 0.05 for all). However cells had a trend of higher ATP production rates and faster proliferation in 5.5 mmol compared to 25 mmol. Fluorescence microscopy revealed the formation of multinucleated myotubes differentiated in 5.5 mmol glucose media containing various concentrations of human serum (0.5%, 1% and 2%).

Conclusions: Our data demonstrates the ability to differentiate LHCN-m2 cells in 5.5 mmol GC, which allows our in vitro model to be more physiologically-relevant and more comparable to what is observed in vivo in humans. Further work is required to determine the implications of GC on the wider metabolic function in LHCN-m2 myoblasts.