Cryopreservation of cells and subcellular fractions

Abstract

The invention provides cryopreserved compositions of cells, wherein the compositions are advantageously in the form of self-sustaining bodies that can be individually handled and combined independently of a container, allowing for easy customization of the eventual pooled preparation. The invention also provides pre-pooled stacks of the self-sustaining cryopreserved compositions for eventual thawing to produce pooled preparations of cells, and methods of creating the same.

What is claimed: 
     
         1 . A combination comprising:
 a first self-sustaining body comprising a first cryopreserved composition of cells; and   a second self-sustaining body comprising a second cryopreserved composition of cells;   wherein said first self-sustaining body and said second self-sustaining body are discrete bodies adjacent one another in a container.   
     
     
         2 . The combination of  claim 1 , wherein said first self-sustaining body and said second self-sustaining body are stacked in physical contact with each other in said container. 
     
     
         3 . The combination of  claim 1 , comprising a plurality of first and second self-sustaining bodies stacked in said container. 
     
     
         4 . The combination of  claim 1 , wherein said first self-sustaining body and said second self-sustaining body can each be handled or moved independently of said container, without being thawed. 
     
     
         5 . The combination of  claim 1 , wherein said first self-sustaining body and said second self-sustaining body each have a thickness of from about 2 mm to about 15 mm, and a width of from about 6 mm to about 20 mm. 
     
     
         6 . The combination of  claim 1 , wherein said first composition comprises cells obtained from a first source and said second composition comprises cells obtained from a second source, said second source being different from said first source. 
     
     
         7 . The combination of  claim 1 , wherein said first composition and second composition each comprise cells obtained from the same source. 
     
     
         8 . The combination of  claim 1 , wherein said cells are hepatocytes. 
     
     
         9 . The combination of  claim 8 , wherein said first and second cryopreserved compositions each comprise from about 1 million viable hepatocytes/mL to about 20 million viable hepatocytes/mL. 
     
     
         10 . The combination of  claim 8 , wherein said hepatocytes are from a single source or from multiple sources. 
     
     
         11 . The combination of  claim 8 , wherein said hepatocytes are human hepatocytes. 
     
     
         12 . The combination of  claim 1 , wherein said cells are cellular or subcellular fractions selected from the group consisting of mitochondria, cytosol, S9, and microsomes. 
     
     
         13 . The combination of  claim 1 , each of said first and second cryopreserved composition of isolated cells being a single-cryopreserved compositions of cells. 
     
     
         14 . The combination of  claim 1 , each of said first and second cryopreserved composition of isolated cells being formed from a suspension of isolated cells dispersed in a solution comprising a culture medium and a cryoprotectant. 
     
     
         15 . The combination of  claim 1 , wherein said container is a vial. 
     
     
         16 . The combination of  claim 1 , each of said first and second cryopreserved compositions further comprising a culture medium and cryoprotectant. 
     
     
         17 . The combination of  claim 16 , wherein said cryoprotectant is selected from the group consisting of fetal bovine serum, dimethyl sulfoxide, polyethylene glycol, amino acids, propanediol, glycerol, and mixtures thereof. 
     
     
         18 . A stack of cryopreserved cells comprising a plurality of discrete, cryopreserved self-sustaining bodies in a container, each of said self-sustaining bodies formed of respective compositions of cells, wherein each of the respective compositions comprises cells from a different source. 
     
     
         19 . A method of creating a stack of cryopreserved cells, said method comprising:
 adding a first self-sustaining body comprising a first cryopreserved composition of cells to a container; and   adding a second self-sustaining body comprising a second cryopreserved composition of cells to said container adjacent to said first self-sustaining body.