Novel Cellular Compositions and Methods for Their Preparation
The present invention relates to novel cell (e.g., hepatocyte, etc.) compositions and methods for their preparation and use. In particular, the invention concerns methods of processing preparations of such cells so as to permit their repeated cryopreservation and thawing while retaining substantial viability. The invention also concerns preparations of cells (e.g., hepatocytes) that have been repeatedly cryopreserved and thawed.
1 . A multi-cryopreserved hepatocyte preparation comprising hepatocytes that have been frozen and thawed at least two times, wherein greater than 50% about 70% of the hepatocytes of said preparation are viable after the final thaw without requiring a density gradient step after thawing the hepatocytes for the second time, wherein the hepatocytes are not plated between the first and second cryopreservations.
2 . The multi-cryopreserved hepatocyte preparation of claim 1 , wherein said hepatocytes are selected from the group consisting of human hepatocytes, porcine hepatocytes, simian hepatocytes, canine hepatocytes, feline hepatocytes, bovine hepatocytes, equine hepatocytes, ovine hepatocytes and rodent hepatocytes.
3 . The multi-cryopreserved hepatocyte preparation of claim 2 , wherein said hepatocytes are human hepatocytes.
4 . The multi-cryopreserved hepatocyte preparation of claim 2 , wherein said preparation comprises a pooled preparation of hepatocytes of multiple sources.
5 . The multi-cryopreserved hepatocyte preparation of claim 4 , wherein said multiple sources are of the same gender, race, or health state.
6 . The multi-cryopreserved hepatocyte preparation of claim 4 , wherein the hepatocytes of said pooled preparation of hepatocytes provide said pooled preparation with a desired level of a metabolic activity.
7 . The multi-cryopreserved hepatocyte preparation of claim 6 , wherein said metabolic activity is selected from the group consisting of coumarin 7-hydroxylase (COUM), dextromethorphan O-demethylase (DEX), 7-ethoxycourmarin O-deethylase (ECOD), activities responsible for the phase II metabolism of 7-hydroxycoumarin (7-HCG and 7-HCS), mephenyloin 4-hydroxylase (MEPH), testosterone 6(β)-hydroxylase (TEST), tolbutamide 4-hydroxylase (TOLB), phenacetin O-deethylase (PHEN), and chlorzoxazone 6-hydroxylase (CZX).
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9 . The multi-cryopreserved hepatocyte preparation of claim 1 , wherein greater than about 80% of the hepatocytes of said preparation are viable.
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21 . The multi-cryopreserved hepatocyte preparation of claim 4 , wherein said multiple sources are of different gender, race, or health state.
22 . The multi-cryopreserved hepatocyte preparation of claim 6 , wherein said multiple sources are selected based upon metabolic activity, and wherein the pooled preparation exhibits a desired level of one or more metabolic activities.