Methods and compositions comprising heat shock proteins
Compositions, and uses thereof, which are beneficial for eukaryotic cells in culture, and methods for their use in promoting cell growth, viability and recombinant protein expression. The methods disclosed in the present application are useful, for example, for improving cell viability and in accelerating the rate of cell growth of cells grown in culture. In one aspect, the supplements of the invention are useful for improving or enhancing the yield of the recombinant proteins from the cell cultures.
1. A cell culture medium supplement composition comprising: a first protein and a second protein, wherein the composition is produced in a plant, wherein said first protein is a protein cell culture component and said second protein is a heat shock protein; and wherein said protein cell culture component comprises at least 0.01% wt/wt of said heat shock protein; and wherein said protein cell culture component has less than 1 EU of endotoxin/mg of said protein cell culture component.
2. The supplement of claim 1 , wherein said heat shock protein is bound to said protein cell culture component.
3. The supplement of claim 2 , wherein said protein cell culture component comprises at least 1% wt/wt of bound long chain fatty acids.
4. The supplement of claim 3 , wherein said supplement provides a cell viability of greater than 10×10 5 cells / m1 AE1 hybridoma cells after 70 hours of culture when said AE1 cells are seeded at an initial density of 0.5×10 5 cells per ml in serum free DMEM with 5 mg/ml supplement and grown under standard culture conditions of humidity and temperature.
5. The supplement of claim 3 , wherein said first protein is albumin.
6. The supplement of claim 5 , wherein said albumin is recombinant albumin.
7. The supplement of claim 6 , wherein said albumin comprises less than 2% wt/wt of aggregated albumin.
8. The supplement of claim 6 , wherein said albumin comprises about 1% to about 2% wt/wt of long chain fatty acids.
9. The supplement of claim 3 , wherein said heat shock protein is a HSP70 family member.
10. A supplement for use in serum free culture, comprising monomeric albumin to which is bound about 1% to about 2% wt/wt of long chain fatty acids and about 0.01% to about 0.1% wt/wt of heat shock proteins.
11. A method for improving the viability of cells in culture comprising the addition of a supplement of any of claims 1 through 3 , or 10 to the cell culture medium.
12. A method for improving the viability of cells grown in culture under serum free conditions, comprising adding the supplement of any one of claims 1 through 3 , or 10 to a cell culture medium used for growing said cells in culture.
13. A method for improving the viability of cells when plated at low density comprising the addition of a supplement of any of claims 1 through 3 , or 10 to the cell culture medium.
14. A method for improving the viability of cells grown in culture from single cell clones comprising the addition of a supplement of any of claims 1 through 3 , or 10 to the cell culture medium.
15. A method for improving the viability of primary cells grown in culture comprising the addition of a supplement of any of claims 1 through 3 , or 10 to the cell culture medium.
16. A method for improving the viability of cells after transfection comprising the addition of a supplement of any of claims 1 through 3 , or 10 to the cell culture medium prior to, during, or immediately after transfection.
17. A method for improving the viability of cells after cryopreservation comprising the addition of a supplement of any of claims 1 through 3 , or 10 to the cell culture medium prior to, during, or immediately after cryopreservation or thawing.
18. A method for improving the yield of a recombinant product produced from cells in culture comprising the addition of a supplement of any of claims 1 through 3 , or 10 to the cell culture medium.
19. The supplement of any one of claims 1 through 3 , or 10 , wherein said first protein is recombinant human serum albumin.
20. The method of claim 12 , wherein the cells are stem cells, or other progenitor cells.
21. The method of claim 17 , wherein the cells are tissue culture cells.
22. The method of claim 18 , wherein the cells are CHO cells.
23. The method of claim 18 , wherein the cells are hybridoma cells.
24. The method of claim 18 , wherein the cells are vero cells or other epithelial cells.
25. The supplement of claim 1 which improves the viability of cells grown in a cell culture medium comprising said supplement.