Pt 5006

hASCs were purchased from Lonza Walkersville, Inc. (Walkersville, MD, USA); the cells were derived from a female donor (non-diabetic, BMI 26 kg/m², 36 years old) (PT5006 lot 0000410257). The cells (passage 4) were pre-cultured as described by Yamada et al. [33 (link)]. Briefly, 2 × 10⁴ cells/mL of the cells were cultured in 50% Dulbecco’s modified Eagle’s medium (DMEM)/50% α minimum essential medium supplemented with 1% fetal bovine serum (FBS), 1 × ITS, 10 ng mL^-1 bFGF (PeproTech Inc., NJ, USA), and 400 ng mL^-1 of hydrocortisone on a 24-well plate for 3 days, until cells became confluent. To induce adipocyte-differentiation, post-confluent preadipocytes were stimulated in MDI differentiation medium (DMEM containing 10% FBS, 1 μM DEX, 0.5 mM IBMX, 0.2 mM indomethacin, 10 μg mL^-1 insulin, and 33 μM biotin) for 7 days (Day 0–7). During cultivation, the culture was replaced with fresh MDI differentiation medium every 2 days with or without Euglena extract. Then, cells were maintained for a further 7 days (Day 7–14) in adipocyte nutrition medium (DMEM containing 10% FBS, 10 μg mL^-1 insulin, and 33 μM biotin), which was replaced with fresh culture every 2 days. All cells were cultured at 37°C under a 5% CO₂ atmosphere. All the above reagents were purchased from Sigma-Aldrich (St. Louis, MO, USA).