The relationship between endogenous oleic acid produced by hepatic delta9-desaturase and the secretion of VLDL-triglycerides was investigated in a primary culture of chicken hepatocytes. When the fatty acid compositions of the secreted and intracellular triglycerides (TG) (or triacylglycerols) were compared, an imbalance between monoenes and saturated fatty acids was observed, with the secreted TG being significantly more unsaturated than the intracellular TG. The addition of a mixture of cyclopropenic fatty acids (specific inhibitors of fatty acid desaturation) to the culture medium of cells 24 h before measurement of their delta9-desaturase activity and TG secretion rate caused a significant impairment of both desaturase activity and TG secretion, without affecting total TG synthesis. However, the addition of oleic acid to the culture medium of cells treated with cyclopropenic fatty acids restored the TG secretion rate. Palmitic acid did not restore the TG secretion rate and linoleic acid partly restored the TG secretion rate. Finally, even in the presence of oleic acid in the culture medium of secreting cells, those which had been treated with cyclopropenic fatty acids had a significantly lower TG secretion rate than nontreated cells. Taken together, these results show that TG secretion is highly dependent on the delta9-desaturase activity and suggest that oversecretion of VLDL-TG in chickens and subsequent fattening could originate in a high hepatic delta9-desaturation of saturated fatty acids.