Previous studies on customizing cell culture environments have utilized a variety of microfabrication-based took to control the spatial localization of adhesive proteins and subsequently mammalian cells. Others have used various methods to immobilize nonadhesive PEO-based polymers on surfaces to inhibit protein absorption and cell adhesion. In this study, we report the application of a well-characterized, commercially available, PEO-terminated triblock polymer (Pluronic(TM) F108) to create micropatterned nonadhesive domains on a variety of biomaterials that deter cell adhesion for up to 4 weeks in culture. The Pluronic can be applied using microfluidic tools or photolithographic techniques, and can be adsorbed to a variety of common surfaces including tissue culture polystyrene, methylated glass, silicone, and polylactic-co-glycolic acid. The effectiveness of the Pluronic in inhibiting cell adhesion in the presence of collagen I is also quantified. Finally, these patterning techniques are generalized to control tissue organization on a variety of common biomaterials. This simple method for micropatterning PEO and, therefore, proteins and cells should prove useful as a tool for biomolecular surface engineering. (C) 2002 John Wiley Sons, Inc.