Adipose-Conditioned Medium Impacts Myogenesis

To generate adipose-conditioned medium (ACM), SAT samples were incubated in differentiation medium at a ratio of 1 g of tissue to 10 mL of medium for 24 h. Larger samples were divided into segments of ~1 g to standardize the surface area of adipose tissue. The ACM was sterile filtered and stored at −80 °C. Prior to use, ACM from numerous patients (normal adiposity: n = 18; excess adiposity: n = 15) was combined to generate a stock of normal adiposity and a stock of excess adiposity ACM, which were matched for mean age and sex distribution (Table 2). The relative abundance of 58 adipokines within both pooled ACM stocks was assessed using a commercially available human adipokine array kit (R&D Systems, #ARY204).
Upon reaching > 90% confluence, myogenic cultures from n = 4 healthy participants (BMI range: 19.8–24.5 kg·m⁻²) were switched to either unconditioned, normal adiposity ACM or excess adiposity ACM for differentiation. Freshly-thawed ACM was diluted 1:2 with fresh differentiation medium and was renewed every 48 h for 6 days [36 (link)].