Foods are not intended to only satisfy hunger and to provide basic nutritional requirements but also to prevent nutrition-related diseases and to improve physical and mental well-being of the consumers. This research studied the improve of nutritional and potential therapeutic value of processed cheese using flaxseed oil because of its high nutritional and health value. Full-fat spreadable processed cheese made by butter substitution levels of nil (control), 25, 50, 75 and 100% with flaxseed oil. Sensory evaluation, chemical, rheological, and biological properties were evaluated to study the effect of addition of flaxseed oil on processed cheese properties. The results of sensory evaluation of samples demonstrated that the best treatment of analogue cheese was the cheese with 25 % of flaxseed oil. As the processed cheese contained flaxseed oil, the total unsaturated increased. The incorporation of the flaxseed oil into the processed cheese caused a high increment in Linoleic, Linolenic and Arachidic acids. The cheese samples with 25 and 50 % of flaxseed oil were the nearest to the control sample towards rheological properties. At the end of the experiment (28 days), rats were fed on processed cheese of flaxseed oil gained higher body weights than those fed only base diet. The total serum cholesterol was lowered in the rats group fed on processed cheese containing 25% flaxseed oil and processed cheese containing 75% flaxseed oil. From these results, it could be recommended flaxseed oil as butter partial substitute in processed cheese manufacture up to 75% to achieve the health benefits propose with good sensory acceptability and reasonable rheological properties.