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Surface properties are largely responsible for the biological performance of biomedical devices, suggesting the great necessity of surface modification. Micro-contact printing (μCP) is a versatile surface modification technique that is capable of not only producing defined topographical features but also manipulating surface chemical and biological cues through customized inks. Compared to other surface patterning techniques, μCP offers distinct advantages of low cost combined with high reliability and versatility. This review summarizes the principles and characteristics of μCP and presents the latest advances enabled by μCP in the biomedical field, categorized by its applications in constructing cell culture platforms, biosensing platforms, and devices for other biological applications.