Milk with an increased content of unsaturated fatty acids was obtained by incorporating 60% of extruded linseed into the concentrate of cows. Two groups of Holstein cows (3 animals/group) were fed a concentrate (control or linseed enriched) together with the same roughage diet (ad libitum). After an adaptation period of 3 wk, evening and morning milk samples were collected every 7 d for 3 wk. Milk was decreamed and anhydrous milk fat (AMF) was isolated from the fat fraction by using the Bureau of Dairy Industries method. The objective of this study was to investigate if the crystallization mechanism of milk fat changed when the content of unsaturated fatty acids was increased. Therefore, the crystallization behavior of a milk fat enriched with unsaturated fatty acids was compared with that of a control milk fat. Nonisothermal crystallization was investigated with differential scanning calorimetry, and 1-step and 2-step isothermal crystallization behaviors were investigated using pulsed nuclear magnetic resonance, differential scanning calorimetry, and x-ray diffraction. A higher content of unsaturated fatty acids in AMF resulted in an increased proportion of low melting triglycerides. These triglycerides lowered the solid fat content profile, particularly at refrigerator temperatures. Furthermore, they induced some changes in the crystallization and melting behaviors of milk fat compared with a control AMF, although no fundamental changes in the crystallization mechanism could be revealed. Even though a lower melting point could be observed for milk fat with a higher content of unsaturated fatty acids, a similar degree of supercooling was needed to initiate crystallization, resulting in a shift in onset temperature of crystallization toward lower temperatures. In addition, slower crystallization kinetics were measured, such as a lower nucleation rate and longer induction times, although crystallization occurred in a similar polymorphic crystal lattice. During melting, a shift in offset temperature toward lower temperatures could be observed for the 3 melting fractions of AMF in addition to a higher proportion of low melting triglycerides. These results demonstrate that a higher content of unsaturated fatty acids has some effect on the crystallization behavior of milk fat. This knowledge could be used to produce dairy products of similar or superior quality compared with conventional products by intervening in the production process of dairy products.