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Cell fate is shaped by external cues and intrinsic cellular states. While supracellular signals, such as growth factors, provide instructive guidance, emerging evidence highlights the role of cell-autonomous properties in modulating these responses. Among these, lipid membrane composition, especially glycosphingolipids (GSLs), has gained attention as a contributor to cellular identity. GSL biophysical properties guide the formation of membrane nanodomains, which serve as platforms for receptor signaling and protein sorting, while their structural heterogeneity enables distinct interactions with signaling molecules and recognition factors. In this review, we discuss the biosynthesis and organization of GSLs, their biological roles, and emerging technologies for their analysis. Together, these studies support the view that GSLs contribute to cell-type-specific membrane properties and help shape cellular states and identity.