Microscale control of cell contact and spacing via three-component surface patterning

The complexity of micropatterned cell constructs has been limited by difficulties in patterning more than two surface components on a culture substrate. Photolithography using multiple aligned masks is well established for generalized multicomponent patterning, but is often too harsh for biomolecules. We report a two-mask photolithographic process that is tuned to preserve bioactivity in patterns composed of covalently coupled poly(ethylene glycol) (PEG), adsorbed extracellular matrix protein (e.g., collagen I), and adsorbed serum proteins (e.g., vitronectin). Thereby, we pattern two cell typesprimary hepatocytes and 3T3 fibroblastsdemonstrating control over contact and spacing (20-200 mu m) between the two cell types for over one week. This method is applicable to the study of intercellular communication in cell biology and tissue engineering.