Tissue scaffolds for electrically excitable cells

Abstract

Inventive concepts relate generally to nanofibrous scaffolds useful for electrophysiological assays. Scaffolds include polymeric nanofibrous components and electrically excitable cells immobilized at a distinct cell seeding domains on the scaffold. Methods and kits including the scaffolds are also described.

1 . A nanofibrous scaffold comprising:
 (a) a layer of unaligned polymeric nanofibers;   (b) a first population of electrically excitable cells immobilized at a first cell seeding domain on the scaffold;   (c) a second population of electrically excitable cells immobilized at a second cell seeding domain on the scaffold at a site that is distinct from the first cell seeding domain; and   (d) at least first and second aligned nanofibrous conduits provided on a surface of the unaligned nanofibers,   wherein the first conduit is in contact with the first cell seeding domain, and the second conduit is in contact with the second cell seeding domain, and   wherein the first conduit and second conduit contact each other at an intersection.   
     
     
         2 . The scaffold according to  claim 1  wherein the unaligned polymeric nanofibers are randomly oriented. 
     
     
         3 . The scaffold according to  claim 1  wherein the electrically excitable cells of the first cell seeding domain are neural cells, and the electrically excitable cells of the second cell seeding domain are selected from neural cells and muscle cells. 
     
     
         4 .- 6 . (canceled) 
     
     
         7 . The scaffold according to  claim 1 , wherein the electrically excitable cells are neural or glial progenitor cells, neural or glial precursor cells, neurotypic cells, pluripotent cells, stem cells, cells which can be differentiated into electrically excitable cells in situ, cells rendered electrically excitable by gene transfection or viral transduction, or any combination of these. 
     
     
         8 . The scaffold according to  claim 1 , wherein the electrically excitable cells are primary cells. 
     
     
         9 . The scaffold according to  claim 1  wherein the electrically excitable cells form synapses at the intersection. 
     
     
         10 . The scaffold according to  claim 1  further comprising at least one electrode operably coupled to scaffold. 
     
     
         11 . The scaffold according to  claim 1  further comprising a support selected from petri dishes, flasks, multiwell plates, slides, films, frames, and hydrogels. 
     
     
         12 . (canceled) 
     
     
         13 . (canceled) 
     
     
         14 . The scaffold according to  claim 1 , wherein the nanofibers comprise at least one chemically reactive functional group or latent reactive group. 
     
     
         15 . The scaffold according to  claim 14 , wherein the latent reactive group comprises anthraquinone, aryl azide, aryl ketone, aryl ketone derivative, halophenyl azide, diazerine, or any mixtures or combinations of any of these. 
     
     
         16 . The scaffold according to  claim 14  wherein the chemically reactive group comprises an amine group. 
     
     
         17 . The scaffold according to  claim 1 , further comprising one or more cell culture agents or gradients of cell culture agents on one or more portions of the scaffold. 
     
     
         18 . (canceled) 
     
     
         19 . The scaffold according to  claim 1  wherein the scaffold includes a microscale or a nanoscale topography that modulates cell growth or proliferation. 
     
     
         20 . The scaffold according to  claim 1  comprising a passivating polymer on a surface of the scaffold. 
     
     
         21 . The scaffold according to  claim 1 , further comprising one or more agents on the aligned conduits to modulate the growth of neurites, axons, dendrites, or their support cells. 
     
     
         22 . The scaffold according to  claim 1  further comprising an electrically conductive material. 
     
     
         23 . The scaffold according to  claim 1  wherein the scaffold comprises a plurality of porous sheets or films assembled in a laminar configuration. 
     
     
         24 . A method of producing a nanofibrous scaffold composite comprising steps of:
 (a) providing a nanofibrous scaffold;   (b) disposing a cell culture agent at two or more distinct domains on a surface of the scaffold, thereby forming two or more distinct cell seeding domains;   (c) disposing two or more aligned nanofibrous conduits on the scaffold surface, in a configuration so that each conduit is in contact with one cell seeding domain, and each conduit intersects the other conduit at an intersection located separate from the cell seeding domains; and   (d) culturing electrically excitable cells on the distinct cell seeding domains under conditions to form functional chemical synapses between the electrically excitable cells.   
     
     
         25 .- 27 . (canceled) 
     
     
         28 . A device for electrophysiological screening, the device comprising:
 (a) a nanofibrous scaffold;   (b) at least first and second cell seeding domains; and   (c) at least first and second nanofibrous conduits arranged on the scaffold, wherein the first conduit is in contact with the first cell seeding domain, and the second conduit is in contact with the second cell seeding domain,   wherein the first conduit and the second conduit contact each other at an intersection, and   wherein the first and second cell seeding domains, and the first and second nanofibrous conduits do not contact each other outside the intersection.   
     
     
         29 . A cell growth substrate comprising:
 (a) a support surface for cellular attachment;   (b) an open field architecture;   (c) wherein an open field potential is generated by a group of electrically excitable cells having been cultured on the substrate;   (d) wherein the generation of an open field potential is enabled by a spatial arrangement of electrically excitable cells, neurites, axons, or dendrites contained in the group of cells; and wherein the arrangement is provided by the open field architecture of the growth substrate.   
     
     
         30 . (canceled)