Animal protein-free media for cultivation of cells

Patent metadata

Field	Value
Publication number	US-10138461-B2
Application number	US-201715712408-A
Country	US
Kind code	B2
Filing date	Sep 22, 2017
Priority date	Oct 29, 2004
Publication date	Nov 27, 2018
Grant date	Nov 27, 2018

Abstract

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.

First claim

What is claimed is:

1. An animal protein-free cell culture medium, comprising at least one polyamine and at least one protein hydrolysate derived from the group consisting of plants and yeast, wherein the polyamine is present in the culture medium in a concentration ranging from about 0.5 to about 30 mg/L.

2. The animal protein-free cell culture medium according to claim 1 , wherein the polyamine is selected from the group consisting of cadaverine, putrescine, spermidine, spermine, agmatine, ornithine, and a combination thereof.

3. The animal protein-free cell culture medium according to claim 1 , wherein the polyamine is putrescine in a concentration ranging from about 0.5 to about 10 mg/L, and the protein hydrolysate is soy hydrolysate in a concentration ranging from about 0.05% (w/v) to about 5% (w/v).

4. The animal protein-free cell culture medium according to claim 1 , wherein the polyamine originates from a source other than a protein hydrolysate.

5. The animal protein-free cell culture medium according to claim 1 , wherein the polyamine is present in the culture medium in a concentration ranging from about 0.5 to about 10 mg/L.

6. The animal protein-free cell culture medium according to claim 1 , wherein the polyamine is present in the culture medium in a concentration ranging from about 2 to about 8 mg/L.

7. The animal protein-free cell culture medium according to claim 1 , wherein the protein hydrolysate is present in the culture medium in a total concentration ranging from about 0.05% (w/v) to about 5% (w/v) for all protein hydrolysates.

8. The animal protein-free cell culture medium according to claim 1 , wherein the protein hydrolysate is derived from a plant selected from the group consisting cereals and soy.

9. A method for cultivating cells, comprising the steps of:
(a) providing an animal protein-free cell culture medium according to claim 1 , and
(b) propagating the cells in the medium to form a cell culture.

10. The method according to claim 9 , wherein the cells are selected from the group consisting of mammalian cells, insect cells, avian cells, bacterial cells, and yeast cells.

11. The method according to claim 9 , wherein the cells are cultivated by a method selected from the group consisting of batch-cultivation, feed-batch-cultivation, perfusion cultivation, and chemostat-cultivation.

12. A method for expressing a target protein, comprising the steps of:
a) providing a culture of cells that have been grown in an animal protein-free cell culture medium according to claim 1 ;
b) introducing a nucleic acid sequence comprising a sequence coding for the target protein into the cells;
c) selecting the cells carrying the nucleic acid sequence; and
d) selectively inducing the expression of the target protein in the cells.

13. The method according to claim 12 , wherein the cells are selected from the group consisting of mammalian cells, insect cells, avian cells, bacterial cells, and yeast cells.

14. The method according to claim 1 wherein the cell/target protein combination is selected from the group consisting of CHO cells/coagulation factor VIII, BHK cells/erythropoietin, Epstein Barr virus transformed, immortalized human B cells/human antibodies.

15. The method according to claim 12 , wherein the cells are cultivated by a method selected from the group consisting of batch-cultivation, feed-batch-cultivation, perfusion cultivation, and chemostat-cultivation.

Assignees

• Baxalta Inc
• Baxalta GmbH

Inventors

• Grillberger Leopold
• Reiter Manfred
• Mundt Wolfgang
• Dorner Friedrich

Classifications

• C12N5/0682

  Cells of the female genital tract, e.g. endometrium; Non-germinal cells from ovaries, e.g. ovarian follicle cells (oocytes C12N5/0609) · CPC title

• C12N2500/92

  Medium free of human- or animal-derived components · CPC title

• C12N2500/76

  from plants · CPC title

• C12N2500/46

  Amines, e.g. putrescine · CPC title

• C12N5/0043 Primary

  Medium free of human- or animal-derived components · CPC title