Apparatus and method for culturing and preserving tissue constructs

Abstract

A disclosure is made of various apparatus and methods for culturing and preserving cells and tissue in ways that minimize contamination potential, direct cells to reside in desired areas, allow uniform cell distribution during seeding, provide optimal growth conditions by controlling the amount of medium residing in proximity of cells, allow desired compounds and molecules to reside in proximity of the cells, allow co-culture, provide for efficient scale up, allow a desired shape of tissue to be created while retaining a closed system, and allow cryopreservation and reconstitution of cell and tissue while retaining a closed system. The apparatus and methods can be combined to prevent the need to remove the tissue from the enclosure at any point during the sterilization, seeding, culturing, cryopreservation, shipping, or restoration process. Also disclosed is an apparatus and method of pipette interface with a container in a manner that blocks contaminants from entering the container.

1 . A cryopreservation and cell culture device including:
 a cryopreservation enclosure, said cryopreservation enclosure being a liquid tight enclosure including at least one access port, and   said cryopreservation enclosure including a tissue bioreactor, and   said tissue bioreactor being a liquid tight container including at least one access port.   
   
   
       2 . The device of  claim 1  wherein said tissue bioreactor is comprised at least in part of gas permeable material. 
   
   
       3 . The device of  claim 1  wherein a gas compartment is defined between an inner surface of the cryopreservation enclosure and an outer surface of said tissue bioreactor. 
   
   
       4 . The device of  claim 3  including a gas compartment access port. 
   
   
       5 . The device of  claim 4  wherein the gas compartment includes a gas filter, one side of said gas filter in contact with ambient gas and the other side of said gas filter in contact with gas residing in said gas compartment. 
   
   
       6 . The device of  claim 3  including a spacer disposed within the gas compartment. 
   
   
       7 . The device of  claim 6  wherein the spacer is a mesh. 
   
   
       8 . A method of cryopreservation using the device of  claim 1 , said method including the steps of:
 adding cells and a cell culture medium into said tissue bioreactor; and   culturing cells; and   removing said cell culture medium and replacing it with a cryopreservation material; and   cryopreserving said cells.   
   
   
       9 . A method of cryopreservation using the device of  claim 1 , said method including the steps of:
 adding cells and a cell culture medium into said tissue bioreactor; and   culturing cells; and   removing said cell culture medium and replacing it with a cryopreservation material; and   adding a cryopreservation material to said gas compartment; and   cryopreserving said cells.