Animal protein free media for cultivation of cells
View Patent ↗The present invention relates to animal protein free cell culture media comprising a combination of non-animal derived peptides derived from soy hydrolysate and yeast hydrolysate. The invention also provides an animal protein free culture process, wherein cells are cultivated, propagated and passaged without animal-derived components. This process is useful for cultivating cells, such as recombinant cells or cells infected with a virus, and for production biological products by cell culture processes under conditions devoid of animal protein components.
1. A method of cultivating mammalian surface-dependent cells to obtain a confluent cell culture, comprising:
providing an animal protein-free medium comprising soy hydrolysate at a concentration of about 0.05% (w/v) to about 1% (w/v) and yeast hydrolysate at a concentration of about 0.05% (w/v) to about 0.3% (w/v);
growing the surface-dependent cells in the medium; and
passaging and sub-cultivating the cells grown in the medium in order to obtain a confluent cell culture;
wherein the cells are infected with an orthomyxovirus and are selected from the grouping consisting of BSC-1 cells, CV-1 cells, COS cells, VERO cells, MDBK cells, MDCK cells, MDOK cells, BHK-21 cells, WI-38 cells, and MRC-5 cells.
2. The method of claim 1 , wherein the cells are recombinant cells.
3. The method of claim 1 , wherein the virus is an influenza virus.
4. The method of claim 1 , wherein the step of passaging and sub-cultivating the cells further comprises contacting the cells with a non-animal protease, wherein the protease is a purified trypsin-like fraction of Strepomyces griseus (SGT).
5. The method of claim 1 , wherein the soy hydrolysate is present in a concentration of between about 0.2% (w/v) to about 0.6% (w/v) and the yeast hydrolysate is present in a concentration of between about 0.05% (w/v) to about 0.2% (w/v).
6. The method of claim 1 , wherein the soy hydrolysate is present in a concentration of between about 0.25% (w/v) to about 0.35% (w/v) and the yeast hydrolysate is present in a concentration of between about 0.05% (w/v) to about 0.15% (w/v).
7. The method of claim 1 , wherein the soy hydrolysate is present in a concentration of about 0.3% (w/v) and the yeast hydrolysate is present in a concentration of about 0.1% (w/v).
8. The method of claim 1 , wherein 3 parts by weight soy hydrolysate are present to 1 part of weight yeast hydrolysate.
9. The method of claim 1 , wherein the yeast hydrolysate is an ultrafiltered purified yeast hydrolysate, and wherein at least 40% of said yeast hydrolysate has a molecular weight of less than or equal to 500 Daltons.
10. The method of claim 1 , wherein the soy hydrolysate is an ultrafiltered purified soy hydrolysate, and wherein at least 40% of said soy hydrolysate has a molecular weight of less than or equal to 500 Daltons.
11. The method of claim 1 , wherein the cells grow on a carrier.
12. The method of claim 10 , wherein the carrier is a synthetic carrier, or a microcarrier coated with a non-animal material.