Four samples of Feta cheese 5 Kg/each made by substituting buffalo milk fat with palm kernel oil (PKO). The substituting ratios were 25%, 50%, 75% and 100% (PKO) to milk fat (MF). Fat content of the obtained cheese was 40% fat/dry matter. A control batch contained milk fat with the same fat content was prepared. The resultant cheese samples were stored at room temperature for three months. Samples were analyzed monthly during pickling period to investigate the effect of the treatment with (PKO) on cheese yield, gross composition, organoleptic properties and the changes in fatty acids pattern of fat content of the resultant cheese samples. The results showed that substituting of milk fat with different ratios of (PKO) caused a significant increase in moisture content, when compared with the control, while the total solids decreased. Cheese yield of all cheese treatments decreased as pickling period progressed. Moisture content of the cheese decreased, whereas fat, total protein and ash contents increased as pickling period progressed. Cheese flavor was enhanced in cheese containing different ratios of (PKO), while the texture of cheese widely depended on the ratio of (PKO) added. Cheese samples showed a similar color and appearance to the control.The changed occurred in fatty acids pattern of the examined samples showed that the addition of (PKO) as a milk fat substitute at ratios 25%, 50%, 75% and 100%, resulted in an increase in the total medium chain fatty acids of the cheese fat when compared with milk fat. Cheese samples treated with 25% and 50% (PKO) were of the higher score of the acceptability due to the presence of medium chain fatty acids. It could be concluded that the use of (PKO) as buffalo's milk fat substitute  succeeded in making Feta cheese of healthy and of acceptance for the consumers..