Apparatus for making engineered tissue using self-assembling cell aggregates

Abstract

A composition comprising a plurality of cell aggregates for use in the production of engineered organotypic tissue by organ printing. A method of making a plurality of cell aggregates comprises centrifuging a cell suspension to form a pellet, extruding the pellet through an orifice, and cutting the extruded pellet into pieces. Apparatus for making cell aggregates comprises an extrusion system and a cutting system. In a method of organ printing, a plurality of cell aggregates are embedded in a polymeric or gel matrix and allowed to fuse to form a desired three-dimensional tissue structure. An intermediate product comprises at least one layer of matrix and a plurality of cell aggregates embedded therein in a predetermined pattern. Modeling methods predict the structural evolution of fusing cell aggregates for combinations of cell type, matrix, and embedding patterns to enable selection of organ printing processes parameters for use in producing an engineered tissue having a desired three-dimensional structure.

1 . (canceled) 
     
     
         2 . A bioprinter cartridge comprising: a storage chamber and a deposition orifice, the storage chamber containing a bio-ink, the bio-ink a solid or semi-solid composition comprising living cells, provided that the bioprinter cartridge is compatible with an computer-aided automatic or semi-automatic bioprinting device. 
     
     
         3 . The bioprinter cartridge of  claim 2 , wherein the bio-ink is extrudable. 
     
     
         4 . The bioprinter cartridge of  claim 2 , wherein the bioprinter cartridge allows the bioprinting device to extrude the bio-ink through the deposition orifice. 
     
     
         5 . The bioprinter cartridge of  claim 2 , wherein the bio-ink is fusible to form a biological structure. 
     
     
         6 . The bioprinter cartridge of  claim 2 , wherein the bio-ink consists essentially of living cells. 
     
     
         7 . The bioprinter cartridge of  claim 2 , wherein the living cells comprise one or more of: muscle cells, liver cells, kidney cells, connective tissue cells, epithelial cells, endothelial cells, neural cells, and precursor cells. 
     
     
         8 . The bioprinter cartridge of  claim 2 , wherein the bio-ink is homocellular. 
     
     
         9 . The bioprinter cartridge of  claim 2 , wherein the bio-ink is heterocellular. 
     
     
         10 . The bioprinter cartridge of  claim 2 , wherein the bio-ink comprises at least three cell types. 
     
     
         11 . The bioprinter cartridge of  claim 2 , wherein the bio-ink is characterized by a high cellular concentration comparable to native mammalian tissues. 
     
     
         12 . The bioprinter cartridge of  claim 2 , wherein the bio-ink comprises one or more extracellular matrix components. 
     
     
         13 . The bioprinter cartridge of  claim 2 , wherein the bio-ink comprises a gel. 
     
     
         14 . The bioprinter cartridge of  claim 13 , wherein the gel is a biodegradable gel. 
     
     
         15 . The bioprinter cartridge of  claim 2 , wherein the bio-ink retains a three-dimensional shape on a flat surface. 
     
     
         16 . The bioprinter cartridge of  claim 2 , wherein a layer of the bio-ink supports the weight of at least one additional layer of bio-ink stacked on top. 
     
     
         17 . The bioprinter cartridge of  claim 2 , wherein the deposition orifice is round. 
     
     
         18 . The bioprinter cartridge of  claim 17 , wherein the deposition orifice has a diameter between 100 and 600 microns.