Cell medium formulation for cell stabilization

Abstract

A cell preservation medium, a cell recovery medium and a cell culture medium, and methods which employ the media, are provided.

1 - 21 . (canceled) 
     
     
         22 . A cell preservation medium comprising: cell hibernation medium and L-glutamine or an analog thereof, and one or more of ascorbic acid, albumin, or insulin. 
     
     
         23 . The medium of  claim 22 , wherein the cell hibernation medium comprises Hibernate® E. 
     
     
         24 . The medium of  claim 22 , wherein the analog comprises L-alanyl-L-glutamine dipeptide. 
     
     
         25 . The medium of  claim 22 , wherein the concentration of the L-glutamine or the analog dipeptide is about 0.5 mM to about 4 mM or about 1 mM to about 2 mM. 
     
     
         26 . The medium of  claim 22 , wherein the ascorbic acid is about 100 μg/mL to about 300 μg/ml or about 150 to about 200 mM. 
     
     
         27 . The medium of  claim 22 , wherein the albumin is about 200 μg/mL to about 700 μg/mL or about 300 to about 400 mM. 
     
     
         28 . The medium of  claim 22 , wherein the insulin is about 1 μg/mL to about 20 μg/mL or about 4 to about 6 mM. 
     
     
         29 . The medium of  claim 22 , further comprising cells. 
     
     
         30 . The medium of  claim 29 , wherein the cells are cardiomyocytes. 
     
     
         31 . The medium of  claim 30 , wherein the cells are iPSc derived cardiomyocytes. 
     
     
         32 . A cell recovery medium comprising: the cell preservation medium of  claim 22 , and M199, DMEM, glucose or other sugar based energy source, glutamine or an analog thereof, and one or more of ascorbic acid, albumin, or insulin, wherein the cell recovery medium has about 45% to about 55% cell preservation medium, wherein the cell recovery medium has about 10% to about 15% M199 and about 35% to about 40% DMEM. 
     
     
         33 . A cell culture medium comprising: M199, DMEM, L-glutamine or an analog thereof, glucose or other sugar based energy source, and one or more of ascorbic acid, albumin or insulin, wherein the medium comprises about 20% to about 30% M199, about 70% to about 80% DMEM, and about 2 g/L to about 8 g/L glucose or other sugar. 
     
     
         34 . The medium of  claim 33  further comprising cells. 
     
     
         35 . The medium of  claim 34  wherein the cells are cardiomyocytes. 
     
     
         36 . The medium of  claim 35  wherein the cells are iPSc derived cardiomyocytes. 
     
     
         37 . A method to prepare cells for storage and shipment, comprising:
 providing cells; and   introducing the cells to the cell preservation medium of  claim 22 .   
     
     
         38 . A method to culture cells, comprising:
 washing cells with the cell recovery medium of  claim 32 ; and   culturing the washed cells with a medium comprising: M199, DMEM, L-glutamine or an analog thereof, glucose or other sugar based energy source, and one or more of ascorbic acid, albumin or insulin, wherein the medium comprises about 20% to about 30% M199, about 70% to about 80% DMEM, and about 2 g/L to about 8 g/L glucose or other sugar.   
     
     
         39 . The method of  claim 38  wherein the cells are cardiomyocytes. 
     
     
         40 . The method of  claim 39  wherein the cells are iPSc derived cardiomyocytes.