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In recent years, new technologies based on 3D bioprinting have emerged as ideal tools with which to arrange cells and biomaterials in three dimensions and so achieve tissue engineering’s original goals. The simplest and most widely used form of bioprinting is based on pneumatic extrusion, where 3D structures are built up by drawing patterns of cell-laden or non–cell-laden material through a robotically manipulated syringe. Developing and characterizing new biomaterials for 3D bioprinting (i.e., bioinks) is critical for the progress of the field. This chapter describes a series of protocols for developing, optimizing, and testing new bioinks for extrusion-based 3D bioprinting.