During adipogenesis, adipocytes accumulate triacylglycerols concomitant with increases in the expression and activity of lipogenic genes such as stearoyl‐CoA desaturase 1 (SCD1). SCD1 is the rate‐limiting enzyme for the conversion of dietary and endogenous saturated fatty acids (SFAs) into monounsaturated fatty acids (MUFAs), specifically, palmitate (PA) to palmitoleate (PMA) and stearate (SA) to oleate (OA). Changes in SCD1 activity alter adipocyte SFA/MUFA ratios, which affects cellular function and the development of obesity‐related complications. We examined the bioactivity of 250µM PA, SA, PMA or OA on SCD1‐inhibited adipocytes. SCD1‐inhibited and differentiated 3T3‐L1 adipocytes were treated with fatty acids (FAs) for 48 hrs. FA composition, gene expression, and cytokine secretion were examined by gas chromatography (GC), qRT‐PCR, and multiplex immunoassays, respectively. Treating SCD1‐inhibited adipocytes with PA caused a substantial increase in SA levels that stemmed from an up‐regulation of elongase 6 activity, which was subsequently confirmed using a PA stable isotope tracer and GC coupled mass spectrometry. Adipocytes treated with SFAs showed larger increases in the expression and secretion of markers of inflammation (MCP1, IL6, and CCL5) compared to MUFAs, and these effects were accentuated with SCD1 inhibition. Taken together, reduced SCD1 activity in adipocytes up‐regulates the conversion of PA into SA, and enhances the pro‐inflammatory nature of SFAs.Grant Funding Source: Supported by NSERC