An arrayed microfluidic platform was developed for cell biology applications. Using soft lithography methods, individual cell culture units were fabricated with a diameter from 50-500 μm. The channel height aspect ratio modulation (CHARM) design allowed fine control of fluidic flow through the array, enabling uniform cell loading, reduced shear stress, and decoupling of fluid transport from cell growth. A variety of adherent mammalian tissue culture cell lines were successfully cultured in the array with continuous perfusion of medium for over 2 weeks. Individual culture units were multiplexed in 2 dimensions to create 6×6 and 8×8 arrays, allowing row and column addressing to provide a unique culture environment in each microchamber. Demonstrated functionalities for cell biology research included repeated cell passaging in a fully enclosed microenvironment, fluorescent cell analysis (live and fixed), on-chip protein tranfection, and multiplexed long-term "flow-cell" microscopy.