Development of 3D Muscle Cell Culture-Based Screening System for Metabolic Syndrome Drug Research

Developing effective drug screening methods for type 2 diabetes requires physiologically relevant models. Traditional 2D cell cultures have limitations in replicating in vivo conditions, leading to challenges in assessing drug efficacy. To overcome these issues, we developed a 3D artificial muscle model that induces insulin resistance, a hallmark of type 2 diabetes. Using C2C12 myoblasts cultured in a scaffold of 1% alginate and 1 mg/mL collagen type 1, we optimized conditions for differentiation and structural stability. Insulin resistance was induced using palmitic acid (PA), and glucose uptake was assessed using the fluorescent glucose analog 2-NBDG. The 3D model demonstrated superior glucose uptake responses compared with 2D cultures, with a threefold increase in insulin-stimulated glucose uptake on days 4 and 8 of differentiation. Induced insulin resistance was observed with 0.1 mM PA, which maintained cell viability and differentiation capacity. The model was validated through comparative drug screening using rosiglitazone and metformin, as well as 165 candidate compounds provided by Korea Chemical Bank. Drug screening revealed that three out of five hit compounds identified in both 2D and 3D models exhibited greater efficacy in 3D cultures, with results consistent with ex vivo assays using mouse soleus muscle. This model closely mimics in vivo conditions, offering a robust platform for type 2 diabetes drug discovery while supporting ethical research practices. Impact Statement The number of patients with type 2 diabetes is increasing every year, and the development of effective drugs is necessary. The shortcomings of 2D cell culture and ethical concerns of animal testing make a need for new drug screening platforms. In this study, we developed an artificial 3D muscle model with induced insulin resistance. This model enabled us to accurately assess the efficacy of drug candidates compared with conventional 2D culture systems. Our findings suggest that this 3D model can serve as a valuable platform for accelerating the discovery of new treatments for type 2 diabetes.