Animal cell culture media comprising peptides derived from rice

• US 6,103,529 A
• Filed: 10/10/1997
• Issued: 08/15/2000
• Est. Priority Date: 10/10/1996
• Status: Expired due to Term

• Current Assignee: Life Technologies Corporation (Thermo Fisher Scientific Incorporated)
• Original Assignee: Life Technologies Incorporated (Thermo Fisher Scientific Incorporated)
• Inventors: Danner, Douglas, Gorfien, Steve, Price, Paul J.
• Primary Examiner(s): Saucier, Sandra E.
• Assistant Examiner(s): Afremova, Vera
• Application Number:
  US08/949,142
• Time in Patent Office: 1,040 Days
• Field of Search: 435/404, 435/390
• US Class Current: 435/404
• CPC Class Codes: C12N 2500/12 Light metals, i.e. alkali, alkaline earth, Be, Al, Mg
• CPC Class Codes: C12N 2500/20 Transition metals
• CPC Class Codes: C12N 2500/22 Zinc; Zn chelators
• CPC Class Codes: C12N 2500/24 Iron; Fe chelators; Transferrin
• CPC Class Codes: C12N 2500/32 Amino acids
• CPC Class Codes: C12N 2500/34 Sugars
• CPC Class Codes: C12N 2500/36 Lipids
• CPC Class Codes: C12N 2500/38 Vitamins
• CPC Class Codes: C12N 2500/74 from fungi, e.g. yeasts
• CPC Class Codes: C12N 5/0043 Medium free of human- or animal-derived components

Abstract

The present invention provides serum-free cell culture media formulations which are capable of supporting the in vitro cultivation of animal cells. The media comprise at least one nutrient of plant derivation, such as at least one plant peptide and/or at least one plant lipid and/or at least one plant fatty acid. The media may further optionally comprise an enzymatic digest or extract of yeast cells. The present invention also provides methods of cultivating animal cells in vitro using these cell culture media formulations. In addition, the media of the present invention can be used for growth of animal cells for virus production.

In particular, an animal cell culture medium comprising at least one peptide derived from rice, wherein said animal cell culture medium is completely devoid of animal-derived proteins.

Claims

1. An animal cell culture medium comprising at least one peptide derived from rice, wherein said animal cell culture medium is completely devoid of animal-derived proteins, and wherein said medium is capable of supporting the cultivation of an animal cell in vitro.

2. The animal cell culture medium of claim 1, said medium further comprising an enzymatic digest or extract of yeast cells.

3. The animal cell culture medium of claim 1 or claim 2, wherein said medium further comprises at least one plant-derived lipid or at least one plant-derived fatty acid.

4. The animal cell culture medium of claim 1 or claim 2, wherein said medium is a 1×

medium formulation.

5. The animal cell culture medium of claim 1 or claim 2, wherein said medium is a 10×

-100×

medium formulation.

6. The animal cell culture medium of claim 1 or claim 2, said medium further comprising at least one ingredient selected from the group of ingredients consisting of at least one amino acid, at least one vitamin, at least one inorganic salt, at least one trace element, at least one plant lipid or fatty acid, adenine sulfate ATP, deoxyribose, ethanolamine, D-glucose, glutathione, N-(2-hydroxyethyl)piperazine-N'"'"'-(2-ethanesulfonic acid) (HEPES), hypoxanthine, linoleic acid, lipoic acid, phenol red, phosphoethanolamine, putrescine, sodium pyruvate, thymidine, uracil and xanthine.

7. The animal cell culture medium of claim 6, wherein said amino acid ingredient comprises one or more amino acids selected from the group consisting of L-alanine, L-arginine, L-asparagine, L-aspartic acid, L-cystine, L-cysteine, L-glutamic acid, glycine, L-histidine, L-isoleucine, L-leucine, L-lysine, L-methionine, L-phenylalanine, L-proline, L-serine, L-threonine, L-tryptophan, L-tyrosine and L-valine.

8. The animal cell culture medium of claim 6, wherein said vitamin ingredient comprises one or more vitamins selected from the group consisting of ascorbic acid magnesium salt, biotin, choline chloride, D-Ca⁺⁺ -pantothenate, folic acid, i-inositol, menadione, niacinamide, nicotinic acid, paraaminobenzoic acid (PABA), pyridoxal, pyridoxine, riboflavin, thiamine, vitamin A acetate, vitamin B₁₂ and vitamin D₂.

9. The animal cell culture medium of claim 6, wherein said inorganic salt ingredient comprises one or more inorganic salts selected from the group consisting of CaCl₂, KCl, MgCl₂, MgSO₄, NaCl, NaHCO₃, Na₂ HPO₄, NaH₂ PO₄ and ferric citrate chelate.

10. The animal cell culture medium of claim 6, wherein said trace element ingredient comprises an ion of one or more trace elements selected from the group consisting of barium, bromium, cobalt, iodine, manganese, chromium, copper, nickel, selenium, vanadium, titanium, germanium, molybdenum, silicon, iron, fluorine, silver, rubidium, tin, zirconium, cadmium, zinc and aluminum.

15. The animal cell culture medium of claim 3, wherein said plant-derived lipid or fatty acid is selected from the group consisting of palmitate, stearate, oleate, linoleate, linolenate, arachidate, myristate, behenate, erucate, lignocerate, caprylate, caprate, laurate and palmitoleate, and combinations thereof.

17. The animal cell culture medium of any one of claims 1 or 11, wherein said animal cell is selected from the group of animal cells consisting of an insect cell, an avian cell, a mammalian cell and a fish cell.

20. A method of cultivating an animal cell comprising the steps of

(a) contacting said animal cell with the animal cell culture medium of any one of claims 1, 2, 11 and 12;

and

(b) cultivating said animal cell under conditions suitable to support cultivation of said animal cell.

21. The method of claim 20, wherein said animal cell is selected from the group of animal cells consisting of an insect cell, an avian cell, a mammalian cell and a fish cell.

22. A composition comprising the animal cell culture medium of any one of claims 1 or 11, and an animal cell.

23. The composition of claim 22, wherein said animal cell is selected from the group of animal cells consisting of an insect cell, an avian cell, a mammalian cell and a fish cell.

24. The composition of claim 22, wherein said mammalian cell is a human cell, a hybridoma cell, a CHO cell, a BHK cell, a COS cell, a VERO cell, a HeLa cell, a 293 cell, a PER-C6 cell, a K562 cell, a MOLT-4 cell, an M1 cell, an NS-1 cell, a COS-7 cell, an MDBK cell, an MDCK cell, an MRC-5 cell, a WI-38 cell, a WEHI cell, an SP2/0 cell, or a derivative thereof.

25. The animal cell culture medium of claim 1 or claim 11, wherein said at least one peptide derived from rice is present at a concentration of about 10 mg/liter to about 1000 mg/liter.

26. The animal cell culture medium of claim 1 or claim 11, wherein said at least one peptide derived from rice is present at a concentration of about 100 mg/liter to about 200 mg/liter.

11. An animal cell culture medium obtained by combining at least one peptide derived from rice together with an animal cell culture medium, wherein said animal cell culture medium is completely devoid of animal-derived proteins, and wherein said medium is capable of supporting the cultivation of an animal cell in vitro.

12. The animal cell culture medium obtained according to claim 11, wherein said medium is obtained by further combining said animal cell culture medium with an enzymatic digest or extract of yeast cells.

13. The animal cell culture medium obtained according to claim 11 or claim 12, wherein said medium is obtained by further combining said animal cell culture medium with a at least one plant-derived lipid or at least one plant-derived fatty acid.

14. The animal cell culture medium obtained according to claim 11, wherein said medium is obtained by combining one or more additional ingredients selected from the group consisting of adenine sulfate, ATP, deoxyribose, ethanolamine•

HCl, D-glucose, glutathione, N-(2-hydroxyethyl)piperazine-N'"'"'-(2-ethanesulfonic acid) (HEPES), hypoxanthine, linoleic acid, lipoic acid, phenol red, phosphocthanolamine, putrescine, sodium pyruvate, thymidine, uracil, xanthine, L-alanine, L-arginine, L-asparagine, L-aspartic acid, L-cystine, L-cysteine, L-glutamic acid, glycine, L-histidine, L-isoleucine, L-leucine, L-lysine, L-methionine, L-phenylalanine, L-proline, L-serine, L-threonine, L-tryptophan, L-tyrosine, L-valine, ascorbic acid magnesium salt, biotin, choline chloride, D-Ca⁺⁺ -pantothenate, folic acid, i-inositol, menadione, niacinamide, nicotinic acid, paraaminobenzoic acid (PABA), pyridoxal, pyridoxine, riboflavin, thiamine, vitamin A acetate, vitamin B₁₂, vitamin D₂, CaCl₂, KCl, MgCl₂, MgSO₄, NaCl, NaHCO₃, Na₂ HPO₄, NaH₂ PO₄, Ba(C₂ H₃ O₂)₂, KBr, CoCl₂, KI, MnCl₂, Cr(SO₄)₃, CuSO₄, NiSO₄, H₂ SeO₃, NaVO₃, TiCl₄, GeO₂, (NH₄)₆ Mo₇ O₂₄, Na₂ SiO₃, FeSO₄, NaF, AgNO₃, RbCl, SnCl₂, ZrOCl₂, CdSO₄, ZnSO₄, Fe(NO₃)₃, AlCl₃ and ferric citrate chelate,

wherein each ingredient is present in an amount which supports the cultivation of an animal cell in vitro.

16. The animal cell culture medium of claim 13, wherein said plant-derived lipid or fatty acid is selected from the group consisting of palmitate, stearate, oleate, linoleate, linolenate, arachidate, myristate, behenate, erucate, lignocerate, caprylate, caprate, laurate and palmitoleate, and combinations thereof.

18. The animal cell culture medium of claim 12, wherein said insect cell is derived from Spodoptera spp. or Trichoplusa spp.

19. The animal cell culture medium of claim 12, wherein said mammalian cell is a human cell, a hybridoma cell, a CHO cell, a BHK cell, a COS cell, a VERO cell, a HeLa cell, a 293 cell, a PER-C6 cell, a K562 cell, a MOLT-4 cell, an M1 cell, an NS-1 cell, a COS-7 cell, an MDBK cell, an MDCK cell, an MRC-5 cell, a WI-38 cell, a WEHI cell, an SP2/0 cell, or a derivative thereof.