The influence of different organic carbon sources (glucose, ethanol, and acetic acid) at different concentrations (0.1, 0.5, and 1.0 g/L for batch and 1.0, 2.0, and 3.0 g/L for fed-batch) were studied in the mixotrophic production (using both light and carbon source) of ${gamma}$-linolenic acid (GLnA) by spirulina (Arthrospira platensis). The obtained spirulina was analyzed in terms of biomass, lipid, and GLnA production. In the batch media, increasing the concentrations of glucose, ethanol, and acetic acid led to an increase in the biomass, lipid, and GLnA production. However, carbon sources at concentrations greater than 1.0 g/L in fed-batch media appeared to have no significant effects on the above parameters. It was also demonstrated that biomass, lipid, and GLnA production using ethanol and acetic acid could be as good as those achieved with the classic glucose-based culture media.