Hagfishes are ancient vertebrates, which have the ability to tolerate nearly a year of food deprivation with energy during fasting maintained using lipid stores. While lipid transporters are evolutionarily conserved, there are relatively few studies examining mechanisms of lipid acquisition in teleosts and no reports in the primitive and evolutionarily important agnathans. We examined tissue lipid droplet distribution and used gut sac preparations to characterize uptake mechanisms of the monounsaturated fatty acid, oleic acid (OA; 18:1 cis-9), in the Pacific hagfish. OA absorption was carrier-mediated and demonstrated saturable concentration-dependent uptake with an affinity constant of 55 ± 7µM, and a maximal rate of uptake of 1311 ± 97pmol cm- 2h- 1. Additionally, regulation of intestinal transport was demonstrated as feeding significantly increased uptake. To further examine post-prandial effects on fatty acid transport, hagfish were dosed with bovine insulin, which had no effect on OA intestinal acquisition, but did lower plasma glucose. Overall, this is the first evidence for intestinal fatty acid acquisition in an agnathan, which are ideal models for understanding the evolution of nutrient transport processes in vertebrates, and are particularly suited to lipid research owing to their dependence upon this energy source during prolonged periods of fasting.