25. Going beyond RGD: screening of a cell-adhesion peptide library in 3D cell culture

Mariah Sarwat, Denver C Surrao, Nick Huettner, James A St John, Tim R Dargaville, Aurelien Forget*

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In tissue engineering, cell-adhesion peptides (CAPs) such as the ubiquitous arginine–glycine–aspartic acid (RGD) sequence have allowed the functionalization of synthetic materials to mimic macromolecules of the extracellular matrix (ECM). However, the variety of ECM macromolecules makes it challenging to reproduce all of the native tissue functions with only a limited variety of CAPs. Screening of libraries of CAPs, analogous to high-throughput drug discovery assays, can help to identify new sequences directing cell organization. However, challenges to this approach include the automation of cell seeding in three dimensions and characterization methods. Here, we report a method for robotically generating a library of 16 CAPs to identify a microenvironment capable of directing a chain-like morphology in olfactory ensheathing cells (OECs), a cell type of particular interest for guiding axon growth in spinal cord injury repair. This approach resulted in the identification of one CAP not previously reported to interact with OECs to direct their morphology into structures suitable for potential axon guidance. The same screening approach should be applicable to any range of cell types to discover new CAPs to direct cell fate or function.