This study aimed to investigate the effects of plant growth regulators in callus cell culture on the oil content and fatty acid composition of the castor bean (Ricinus communis L.). Ricinus communis L. are valued for its oil and the performance of oil is closely related to its fatty acid composition. Thus, producing oil in vitro with favored fatty acid profiles is a promising research area and may also offer industrial opportunities. In line with this, the total amount of fat and the fatty acid composition of the samples, which were endosperm and calli obtained by treatment of various doses of plant growth regulators were determined. Results showed that the type and amount of the plant growth regulator used in the media affect the fatty acid composition. In detail, the biggest change was shown by Indole-3-Acetic Acid (IAA), in general, using the plant growth regulators at 5 mgL-1, instead of 20 mgL-1, was found to have induced larger differentiations. The effect of a natural plant growth regulator (IAA) on fatty acid profiles was larger than the synthetic ones (NAA, 1-Naphthaleneacetic acid, and 2,4 D, 2,4-Dichlorophenoxyacetic acid). The media containing 5 mgL-1 of NAA, 20 mgL-1 of NAA, 20 mgL-1 of 2,4 D, or 5 mgL-1 of 2,4 D gave similar results.