Tissue scaffolds for electrically excitable cells

A method of producing an electrophysiological assay scaffold comprising steps of:

• (a) providing a nanofibrous scaffold composed of unaligned polymeric nanofibers that form a nanofiber support having a planar surface;
• (b) printing, spraying, or brushing a cell culture agent at two or more physically isolated domains on the planar surface, thereby forming at least a first cell seeding domain and a second cell seeding domain, wherein the cell culture agent is present only at the cell seeding domains;
• (c) laminating the nanofiber support with a first planar-aligned nanofiber assembly and a second planar-aligned nanofiber assembly by placing the first planar-aligned nanofiber assembly at a location on the planar surface such that the first planar-aligned nanofiber assembly contacts the first cell seeding domain and does not contact the second cell seeding domain, and placing the second planar-aligned nanofiber assembly at a location on the planar surface such that the second planar-aligned nanofiber assembly contacts the second cell seeding domain, does not contact the first cell seeding domain, and contacts the first planar-aligned nanofiber assembly at an intersection that is located separate from the cell seeding domains, thereby forming a laminated scaffold;
• (d) annealing the first planar-aligned nanofiber assembly and second planar-aligned nanofiber assembly to the planar surface;
• and
• (e) culturing neurons on the distinct cell seeding domains under conditions to form functional chemical synapses between the neurons of the distinct cell seeding domains.