Diverse external stimuli such as surrounding matrix proteins, neighboring cells, and soluble factors define state of cell differentiation or proliferation. Our research focuses on developing intelligent surfaces to enable parallel investigation of multiple external stimuli in order to expedite discovery of inducers of tissue-specific differentiation. For this purpose, photolithography and protein microarraying are combined to create complex micropatterns that represent multiple microenvironment scenarios. Silane-modified glass substrates were first patterned with photoresist to create periodic patterns of 60, 40, 20, 10 µm squares. In a second step, protein microarrays containing extracellular matrix (ECM) proteins (collagen type I, IV, laminin, and fibronectin) and poly-L-lysine were printed over the photoresist pattern. The photoresist served as a stencil during arraying and defines micrometer-scale cell-adhesion domains within each spot. After removal of the photoresist in organic solvents, each printed protein spot (500 µm or 150 µm in diameter) contained clusters of micrometer-scale cell-adhesive regions. Hepatocytes seeded on these micropatterned substrates attached to different ECM components and form single cell patterns as well as multi-cell clusters of varying sizes signifying different cell-cell interaction scenarios. Therefore, each microfabricated substrate creates combination of microenvironment conditions with cell-substrate and intercellular interactions being varied in a location specific fashion along the substrate. Methods for creating complex, combinatorial cellular microenvironment need to be coupled to novel techniques for analysis of cell function in the context of local microenvironment. This presentation will also describe our efforts to selectively retrieve small populations of hepatocytes from micropatterned surfaces using laser microdissection for downstream analysis of liver-specific gene expression by quantitative real-time RT-PCR. Novel methods for cell culture and analysis discussed here will be particularly useful for culturing scarce cells (e.g. human hepatocytes) or for discovery of inducers of stem cell differentiation.