A Systematic Review and User Reference of Phenotypic and Molecular Characteristics of Dexamethasone‐Mediated C2C12 Muscle Atrophy

BACKGROUND: Skeletal muscle is a vital part of human physiology and is responsible for numerous essential functions. Not surprisingly, the loss of skeletal muscle mass and function is common in several pathologies including atrophy and sarcopenia, which profoundly impact quality of life of those afflicted. Thus, numerous investigations of potential therapies for mitigating or reversing such pathologies are available. Within these studies, experimental cell culture models such as the murine C2C12 myoblasts are commonly used. Over 100 publications have utilized dexamethasone-treated C2C12 myotubes to investigate various aspects of muscle atrophy. The purpose of this systematic review is to describe the experimental conditions common to these experiments, as well as phenotypical myotube presentation, and gene and protein expression of targets that regulate muscle mass, function, and metabolism. METHODS: A systematic review of literature was conducted until 3 January 2025 using PUBMED. Articles were included if (1) C2C12 myotubes were used, (2) the article included a dexamethasone-only group along with appropriate vehicle or true control and (3) the article assessed at least one of the related phenotypical or molecular outcomes of importance to the scope of the review. RESULTS: A total of 182 articles were included after screening for relevance and inclusion criteria, which were assessed for outcomes (raw data reported when available or using ratio-metric estimates of relative differences between dexamethasone treatment and control). In 24 of 26 unique experiments that utilized 10 μM dexamethasone and 37 of 39 unique experiments that utilized 100 μM dexamethasone, a decrease in myotube diameter was reported (pooled experimental average estimates from 24-h time points 69.8% ± 7.5% and 66.9% ± 14.7% for 10 and 100 μM, respectively, vs. control). All six studies that utilized 10 μM dexamethasone and all nine that treated myotubes with 100 μM dexamethasone reported reduced fusion index (pooled experimental average estimates from 24-h time points: 67.6% ± 5.3% and 68.4% ± 8.4% for 10 and 100 μM, respectively, vs. control). Dexamethasone-treated myotubes also consistently expressed increased atrophic-related molecular targets including Atrogin-1 and muscle atrophy X box 1 (MuRF1), as well as reductions in anabolic signalling (specifically, mTORC and Akt activation) and mitochondrial function. CONCLUSIONS: The striking consistency of these findings suggests dexamethasone treatment of C2C12 myotubes is a reliable method of mimicking many features common to skeletal muscle pathology. This review provides insight into the use and expected outcomes of the dexamethasone-mediated model of atrophy in C2C12 myotubes and may serve as a helpful reference for future experiments utilizing this model.