Guava (Psidium guajava) is a widely consumed fruit and has been commercialized in markets. The seeds are by-products of the processing procedures performed by the commercial guava juice industry. They are considered a nutritional resource that has been poorly utilized as they contain essential fatty acids such as linoleic acid (LA) and phenolics in abundance. In the study, guava seed oil (GSO) was used, which was obtained by hexane extraction of guava seeds to determine composition and test toxicity, cell migration, cancer cell viability, and plasmodium growth. GSO was found to be relatively nontoxic to normal hepatocytes and peripheral blood mononuclear cells, with mice for 14 days showing median lethal dose (LD 50 ) > 10 mg/kg and rats for up to 90 days. Surprisingly, the oil inhibited the proliferation of the human erythroleukemic cells in a dose-dependent manner with the half maximal inhibitory concentration values of 155 and 137 μg/ml at 24 and 48 hr, respectively. Importantly, GSO at 500 μg/ml was found to increase the degree of migration of keratinocytes (HaCaT). These observations suggest that edible P. guajava seed oil, which is abundant with linoleic acid and antioxidants, can promote skin wound healing and inhibit the proliferation of leukemic cells.