On the basis of earlier reports of reduced growth rate and fat accumulation in animals fed trans 18:1, a study was conducted to determine whether the effects of trans 18:1 on lipolysis and glucose utilization by adipocytes differed from effects of the cis isomer. Two experiments compared three fatty acid isomers (oleic, elaidic and vaccenic acids) at several concentrations and at several fatty acid to albumin ratios in cell media. Adipocytes were isolated from adipose tissue of rats by collagenase digestion and incubated for 2 h in media containing added fatty acids. Compared with oleic acid, both trans isomers reduced (P < 0.01) the amount of glucose converted to cell lipid in both experiments. Glucose oxidation to carbon dioxide also was lower for both trans fatty acids in Experiments 1 (P < 0.05) and 2 (P < 0.06). Lipolytic rates were increased (P < 0.01) in both experiments by replacing oleic acid with either of the trans isomers. Trans isomers of octadecenoic acid had catabolic effects on adipocyte metabolism that occurred regardless of the position of the double bond, the fatty acid concentration in media or the fatty acid to albumin ratio. These catabolic effects explain previous observations of reduced growth rate and fat accumulation when oleic acid in animal diets is replaced with a trans isomer.