Animal protein-free media for cultivation of cells
View Patent ↗The present invention relates to animal protein-free cell culture media comprising polyamines and a plant- and/or yeast-derived hydrolysate. The invention also relates to animal protein-free culturing processes, wherein cells can be cultivated, propagated and passaged without adding supplementary animal proteins in the culture medium. These processes are useful in cultivating cells, such as recombinant cells or cells infected with a virus, and for producing biological products by cell culture processes.
1. A method for cultivating Chinese hamster ovary (CHO) cells in soy hydrolysate-containing animal protein-free medium supplemented with putrescine to overcome inhibitory effects on cell growth due to soy hydrolysate lot variation, said method comprising the steps of:
(a) providing an animal protein-free cell culture medium comprising soy hydrolysate and putrescine, wherein the putrescine is added to said culture medium in a concentration ranging from 0.5 mg/L to 10 mg/L, and the soy hydrolysate is present in a concentration ranging from 0.05% (w/v) to 0.5% (w/v); and
(b) propagating the cells in the medium to form a cell culture.
2. The method according to claim 1 , wherein the CHO cells are cultivated by a method selected from the group consisting of batch-cultivation, feed-batch-cultivation, perfusion cultivation, and chemostat-cultivation.
3. The method according to claim 1 , wherein the CHO cells comprise a nucleic acid comprising a sequence encoding a target protein, and the CHO cells are capable of expressing said target protein.
4. The method according to claim 3 , wherein the target protein is at least a biologically active part of a blood coagulation factor, or at least a biologically active part of a protein involved in the production of red blood cells and angiogenesis, or a monoclonal antibody.
5. The method according to claim 4 , wherein the target protein is Factor VIII or erythropoietin.
6. The method according to claim 3 , wherein the cells express said target protein, and the method further comprises the step of isolating the target protein.
7. The method according to claim 1 , wherein the putrescine is added in a concentration from 2 mg/L to 8 mg/L.