The fat in breast milk exists in the form of fat globules, which are enveloped by a unique biophysical membrane called the milk fat globule membrane. The absence of fat globules will affect the structure and stability of fat droplets, which will in turn affect lipid digestion and absorption. In an effort to make the nutritional composition of milk powder approximate as closely as possible to that of breast milk, an extensive literature has developed concerning the fat globule membrane of cow’s milk, goat’s milk and other milks both domestically and internationally. With the aim of providing a reference for the resource development and application of milk fat globule membrane resources, this article reviews recent progress in understanding the composition of the milk fat globule membrane as well as the factors affecting the structure and functional characteristics of the milk fat globule membrane such as lactation stages, feeding regimes and milk sources as well as separation, thermal processing, homogenization and non-thermal processing, and it summarizes recent advances in breast milk fat globule membrane simulation and its application in infant formula and milk products for middle-aged and elderly people.