Materials

Adipose derived-stem cells (ADSCs) were purchased directly from Lonza (Poietics™ Normal ADSCs, Lonza, PT-5006, Lot 0F4505, Switzerland). These commercially available ADSCs are isolated from normal (non-diabetic) adult subcutaneous lipoaspirates collected during elective surgical liposuction procedures. ADSCs have been reported to differentiate into many different lineages, including chondrogenic, osteogenic, adipogenic and neural lineages. Adipogenesis media and reagents were obtained from Lonza, and 3-AT was purchased from Sigma (Sigma-Aldrich, St. Louis, MO, USA). Adenosine 3′,5′-cyclic monophosphate (cAMP) was acquired from Sigma (A9501). The rabbit anti-GLUT4 antibody (H-61) and TNF-α antibody (SC-52746) were acquired from Santa Cruz Biotechnology (Santa Cruz, CA, USA). The goat anti-adiponectin antibody (AF1065) was obtained from R&D Systems (R&D, Inc. USA). The rabbit anti-PPARγ (D69), phosphor-NF-kB p65 (Ser536), rabbit anti-total AMPKα (D5A2) and rabbit anti-phospho-AMPKα (Thr172) antibodies were acquired from Cell Signaling Technologies (Beverly, MA, USA). The mouse anti-α-tubulin antibody (T5158) and horseradish peroxidase-conjugated immunoglobulin were purchased from Sigma. Unless otherwise indicated, all other chemicals were purchased from Sigma.