Biomaterials are increasingly being developed as in vitro microenvironments mimicking in vivo stem cell niches. However, current macroscale methodologies to produce these niche models fail to recapitulate the spatial and temporal characteristics of the complex native stem cell regulatory systems. Microfluidic technology offers unprecedented control over the spatial and temporal display of biological signals and therefore promises new avenues for stem cell niche engineering. Here we discuss how the two approaches can be combined to generate more physiological models of stem cell niches that could facilitate the identification of new mechanisms of stem cell regulation, profoundly impacting drug discovery and ultimately therapeutic applications of stem cells.