We want to study senescence in adipocytes. However, we applied the method at adipose cells in our laboratory was as follows; ''Preadipocyte cell line 3T3-L1 (American Type Culture Collection, Manassas, VA, USA, CL173™) were cultured in low glucose (1 g/L) DMEM supplemented with 10% calf serum, 100 g/mL streptomycin, and 100 U/mL penicillin. This is known as complete medium. The cells were grown in culture flasks and the complete medium was changed every 2-3 days and incubated at 37°C in a humidified environment (%5 CO2, 95% air). Differentiation of the 3T3-L1 cell line Differentiation of 3T3-L1 preadipocytes was performed according to standard protocols (Green and Kehinde 1975). As a brief description, the preadipocytes were seeded in 6-well plates with complete medium and left until confluence. Next, the differentiation process was induced by incubating with differentiation medium I (DMI), which contains high glucose DMEM (4,5 g/L) supplemented with 10% FBS and adipogenic reagents such as 10 μg/mL insulin, 1 μM DXMT and 0.5 mM IBMX. This DMI was left for 48 hours and then changed to differentiation medium II (DMII), which contains insulin and 10% FBS in high glucose DMEM (4,5 g/L). The DMII was changed every 2-3 days for 10-12 days, obtaining mature adipocytes. Hypertrophic adipocyte model Once the differentiation to mature adipocytes has occurred, the hypertrophic state of the adipocytes can be obtained in order to simulate obese adipose tissue by incubating with high glucose concentrations for a long period. To obtain hypertrophic adipocytes from mature adipocytes, the cells were maintained with DMII for a minimum of 7 additional days. Nearly after 20 days of incubation, a hypertrophic adipocyte model has been observed. It is a well-established model of insulin resistance in adipocytes subjected to metabolic stress '' We do not yet have any information about aging processes. Thank you very much in advance for your sharing. best regards sincerely, Associate Professor Emine Kilic-Toprak