Preclinical experimentation demands for highly reliable and physiologically-relevant systems capable of recapitulating the complex human physiology. Further technological advances are in great need for improving our understanding about critical biological processes involved in tissue development or cancer progression, and for the discovery and screening of novel pharmacological drugs. Traditional in vitro models, albeit widely employed, fail to reproduce the complexity of the native scenario. Similarly, in vivo animal models poorly mimic the human condition and they are ethically questionable. During the last two decades, a new paradigm in preclinical modelling has emerged aiming to solve the limitations of the previous methods. The combination of advanced tissue engineering, cell biology and nanotechnology, has resulted in the development of cutting-edge microfluidics-based models with an unprecedented ability to recreate the native habitat of cells within a microengineered chip. Among the diverse variety of micro- and bio-fabrication techniques, UV-photolithography and soft lithography are considered the gold-standard methods for the fabrication of microfluidic chips to their simplicity, versatility, and rapid prototyping. In this paper, we describe a protocol for the fabrication of a microfluidic chip by UV-photolithography and replica molding, and an example of its use in cell migration assays.