Three-dimensional culture device and methods for dynamic culture of cell aggregates

Abstract

The subject invention concerns materials and methods for culture of cell aggregates. The subject invention utilizes three-dimensional (3-D) inserts comprising micro-channels having selected dimensions. The inserts are provided in or on tissue culture plates that can be supported on a programmable rocking platform/station, thereby providing for a hydrodynamic environment that promotes 3-D aggregation of cells cultured on the plates. In one embodiment, one or more micro-channels are provided in a 3-D insert that can be provided in or placed within the tissue culture plate. Cells are placed in the micro-channels of the inserts in the tissue culture plates that comprise a suitable cell growth medium for the particular type of cell. The supporting rocker is programmed to provide motion that generates hydrodynamic conditions that support 3-D cell aggregation and long-term culture. Aggregated cells can be harvested and isolated from the micro-channels of the 3-D inserts in the tissue culture plate at suitable times. The subject invention also concerns an apparatus comprising a tissue culture vessel that comprises a 3-D insert of the present invention, and a programmable rocking platform/station that can provide motion to the vessel provided thereon, thereby generating hydrodynamic conditions and wave motion that support 3-D cell aggregation and cell culture. The subject invention also concerns methods for growing 3-D aggregates of cells. Methods of the invention comprise culturing cells in a micro-channel of a 3-D insert of the invention provided in or as part of a tissue culture vessel or the well of a tissue culture vessel.

What is claimed is: 
     
         1 . A 3-D tissue culture insert comprising one or more micro-channels that provide for a confined space for cells that are cultured in said micro-channels. 
     
     
         2 . The insert according to  claim 1 , wherein said micro-channels have a width of about 1 mm to about 5 mm, a length of about 20 mm to about 60 mm, and a height of about 5 mm to about 15 mm. 
     
     
         3 . The insert according to  claim 1 , wherein the insert fits into the well of a tissue culture vessel. 
     
     
         4 . The insert according to  claim 1 , wherein one or more surfaces of said insert is coated with or composed of a material that inhibits or prevents adherence or attachment of a cell to said surface. 
     
     
         5 . A method for growing cells into aggregates of cells, the method comprising culturing a cell in a micro-channel of a 3-D insert of  claim 1  provided in a tissue culture vessel, wherein said vessel comprises suitable culture media for culturing said cell. 
     
     
         6 . The method according to  claim 5 , wherein said cell is a mammalian cell or a human cell. 
     
     
         7 . The method according to  claim 5 , wherein said cell is a stem cell. 
     
     
         8 . The method according to  claim 7 , wherein said stem cell is a mesenchymal stem cell. 
     
     
         9 . The method according to  claim 5 , wherein said micro-channel has a width of about 1 mm to about 5 mm, a length of about 20 mm to about 60 mm, and a height of about 5 mm to about 15 mm. 
     
     
         10 . The method according to  claim 5 , wherein one or more surfaces of said insert is coated with or composed of a material that inhibits or prevents adherence or attachment of a cell to said surface. 
     
     
         11 . The method according to  claim 5 , wherein the method further comprises rocking said vessel at a frequency to generate hydrodynamic conditions to support cell aggregation. 
     
     
         12 . The method according to  claim 11 , wherein said rocking is at a frequency of about 1 to 5 rocks per minute. 
     
     
         13 . The method according to  claim 11 , wherein said rocking is at a rocking angle of about 15 to about 20 degrees. 
     
     
         14 . The method according to  claim 5 , wherein the method further comprises harvesting said cell aggregates after about 18 to 120 hours of culturing said cell in said micro-channel. 
     
     
         15 . The method according to  claim 14 , wherein said harvested cells are seeded back into culture media for further culturing. 
     
     
         16 . The method according to  claim 5 , wherein said insert is composed of a plastic (such as polystyrene, polypropylene, or polycarbonate), glass, ceramic, metal, metal alloy, or composite. 
     
     
         17 . The method according to  claim 5 , wherein said insert is round, square, triangular, rectangular, pentagonal, hexagonal, heptagonal, octagonal, nonagonal, or decagonal in shape. 
     
     
         18 . The method according to  claim 5 , wherein said insert is removable or detachable from said tissue culture vessel. 
     
     
         19 . A tissue culture vessel comprising a 3-D insert of  claim 1 ; or
 an apparatus comprising said tissue culture vessel and a programmable rocking platform or station that can provide motion to said vessel to thereby generate hydrodynamic conditions to support 3-D cell aggregation.   
     
     
         20 . The tissue culture vessel according to  claim 19 , wherein said vessel comprises tissue culture media that supports the growth of cells.