Cell substrate patterning with glycosaminoglycans to study their biological roles in the central nervous system.

Microcontact printing (μCP) based techniques have been developed for creating cell culture substrates with discrete placement of CNS-expressed molecules. These substrates can be used to study various components of the complex molecular environment in the central nervous system (CNS) and related cellular responses. Macromolecules such as glycosaminoglycans (GAGs), proteoglycans (PGs), or proteins are amenable to printing. Detailed protocols for both adsorption based as well as covalent reaction printing of cell culture substrates are provided. By utilizing a modified light microscope, precise placement of two or more types of macromolecules by sequential μCP can be used to create desired spatial arrangements containing multicomponent PG, GAG, and protein surface patterns for studying CNS cell behavior. Examples of GAG stripe assays for neuronal pathfinding and directed outgrowth, and dot gradients of PG + laminin for astrocyte migration studies are provided.