Cell culture media containing combinations of proteins
The present invention relates to cell culture media supplemented with a plant-produced recombinant mammalian albumin supplement, as well as methods of making the cell culture media, and methods of using the supplemented cell culture media to improve growth characteristics of cultured cells.
1. A method for enhancing cell growth in culture, said method comprising:
a) culturing cells in a cell culture media supplemented with a plant-produced recombinant mammalian albumin supplement, wherein said plant-produced recombinant mammalian albumin has less than 1 EU of endotoxin/mg of albumin and less than 2% aggregated albumin, wherein said cell growth in culture is enhanced relative to a culture without said plant-produced recombinant mammalian albumin supplement.
2. The method of claim 1 , wherein said cell culture media is fetal bovine serum-free media.
3. The method of claim 1 , wherein said albumin is recombinant human serum albumin.
4. The method of claim 1 , wherein said plant-produced recombinant mammalian albumin supplement is a transgenic grain extract.
5. The method of claim 4 , wherein the transgenic grain is transgenic rice.
6. The method of claim 1 , wherein said plant-produced recombinant mammalian albumin supplement is added to said cell culture media at day 0 of said culturing.
7. The method of claim 1 , wherein said plant-produced recombinant mammalian albumin supplement is added to said cell culture media before day 4 of said culturing.
8. The method of claim 1 , wherein said plant-produced recombinant mammalian albumin supplement is added to said cell culture media daily during said culturing.
9. The method of claim 1 , wherein said recombinant mammalian albumin comprises at least 0.01% wt/wt of a heat shock protein.
10. The method of claim 9 , wherein said heat shock protein is a rice heat shock protein.
11. The method of claim 10 , wherein said heat shock protein is selected from the group consisting of Rice HSP70 proteins, and homologs thereof selected from the group consisting of rice endosperm lumenal binding protein (Bip homolog) and rice stromal protein.
12. The method of claim 11 , wherein said supplement comprises at least 0.04% wt/wt HSP70.
13. The method of claim 10 , wherein said heat shock protein is selected from the group consisting of Rice (gb|ACJ54890.11), EEC69073/OsI_37938, and AAB63469.
14. The method of claim 1 , further comprising a recombinant transferrin related protein, wherein said transferrin related protein is lactoferrin.
15. The method of claim 14 , wherein said ratio of said lactoferrin to said recombinant mammalian albumin is 1:3 to 1:0.33.
16. The method of claim 1 , further comprising a recombinant transferrin related protein, wherein said transferrin related protein is transferrin.
17. The method of claim 16 , wherein said ratio of said transferrin to said recombinant mammalian albumin is 1:50 to 1:5000.
18. The method of claim 3 , wherein said plant-produced recombinant human serum albumin is an expression product of a transgenic monocot plant.
19. The method of claim 18 , wherein said transgenic monocot plant is a transgenic rice plant ( Oryza sativa ).
20. The method of claim 19 , wherein said plant-produced recombinant human serum albumin is a seed protein extract from a rice seed of said transgenic rice plant.
21. The method of claim 19 , wherein said cell culture media is further supplemented with rice heat shock protein.
22. The method of claim 21 , wherein said rice heat shock protein is a seed protein extract from a rice seed of said transgenic rice plant.
23. The method of claim 21 , wherein said rice heat shock protein is co-purified from said transgenic rice plant with said plant-produced recombinant human serum albumin.
24. The method of claim 18 , wherein said plant-produced recombinant human serum albumin has a purity of greater than about 95% as measured by SDS-PAGE.
25. The method of claim 1 , wherein said cells are selected from the group consisting of tissue culture cells, CHO cells, hybridoma cells, VERO cells, stem cells, B-cells, T-cells, B-cell derived cells, and T-cell derived cells.
26. The method of claim 1 , further comprising culturing said cells having enhanced growth for in vitro fertilization, testing an effect of a compound or drug on said cells, synthesizing biologics, propagation and differentiation of stem cells, vaccine production, or tissue engineering.