This study aimed to investigate the utilization of cold-pressed chia-seed oil by-products (CSOB) in a low-fat ice cream formulation as a fat replacer and stabilizer. In the study, ice cream emulsion mixtures were formulated by using 0.2-0.4% xanthan gum (XG), 2.5-12.5% fat, and 1-3% CSOB. Optimization was performed using the response surface methodology (RSM) and full factorial central composite design (CCD) based on the flow behavior rheological properties of the emulsions obtained from 17 different experimental points. All of the emulsion samples showed non-Newtonian shear-thinning flow behavior. The consistency coefficient (& UKappa;) values of the emulsion samples were found to be 4.01-26.05 Pas(n) and were significantly affected by optimization parameters (p < 0.05). The optimum formulation was determined as 0.29% XG, 2.5% CSOB, 2.5% fat. The low-fat (LF-IC) and full-fat control samples (FF-IC) were compared to samples produced with an optimum formulation (CBLF-IC) based on the steady shear, frequency sweep, and 3-ITT (three interval thixotropy test) rheological properties, thermal properties, emulsion stability, light microscope images, and sensory quality. CBLF-IC showed similar rheological behavior to FF-IC. The mix of CBLF-IC showed higher emulsion stability and lower poly-dispersity index (PDI) value and fat globule diameters than those of FF-IC and LF-IC. The thermal properties of the samples were significantly affected by the addition of CSOB in an ice cream mix. CBLF-IC exhibited a lower temperature range (& UDelta;T), enthalpy of fusion (& UDelta;H-f), and freezing point temperature (T-f) than those of FF-IC and LF-IC. While CBLF-IC exhibited a higher overrun value than other samples, it showed similar sensory properties to the FF-IC sample. The results of this study suggested that CSOB could be used successfully in low-fat ice cream production. This study also has the potential to gain new perspectives for the evaluation of CSOB as a fat substitute in a low-fat ice cream.