The immortal subcutaneous human normal preadipocyte (NPAD) clone B cell line has been described previously (Nitta et al. 2013 (link), Vu et al. 2013 (link), Gadupudi et al. 2015 (link), Littlejohn et al. 2016 (link)). Cells were cultured and passaged as preadipocytes in preadipocyte growth media, PGM2, a proprietary rich media with high buffering capacity supplemented with glutamine (2mM), and 10% FBS, along with gentamycin (30 μg/ml) and Fungizone (15 ng/ml) according to the manufacturer’s recommendations (Lonza). For differentiation, cells were plated in either 6-well plates or 35 mm dishes at 80,000 cells/well or dish and allowed to grow to confluency for 4 days before addition of differentiation media, PDM2 (Lonza). Preadipocyte differentiation media, PDM2, is PGM2 media with added differentiation components from a Bullet Kit (Lonza) including dexamethasone (final concentration of 1 μM, IBMX (final concentration of 0.5 mM), insulin (final concentration of 15 μg/ml), and indomethacin (final concentration of 0.2 mM), all according to the manufacturer’s instructions. For differentiation, cells were kept in PDM2 media for 10–14 days as recommended by Lonza and as described previously (Gadupudi et al. 2015 (link)) with a media change (PDM2) at 5 days.