Milk protein mixture for promoting growth of animal cells or tre

Details Milk protein mixture for promoting growth of animal cells or tre Milk protein mixture for promoting growth of animal cells or tre

1992-12-07

1999-02-02

Sayala, Chhaya D.

Chemistry: molecular biology and microbiology

Animal cell, per se ; composition thereof; process of...

Method of culturing cells in suspension

435325, 424535, 530350, 530399, 530416, C12N 506

Patent

active

058664181

DESCRIPTION:

BRIEF SUMMARY
This invention relates to the growth of animal cells in a cell culture composition. More specifically it relates to the provision of a cell culture composition including a cheese whey extract composition.
Animal cells are grown in culture to provide a number of pharmaceutical, diagnostic and veterinary products including human vaccines, lymphokines, hormones, monoclonal antibodies, Other pharmaceutically active protein products, and veterinary hormones and for research and development and diagnostic purposes.
The growth of animal cells requires a defined isotonic medium that contains salts, nutrients, lipid precursors, nucleic acid precursors, vitamins and amino acids that are formulated to mimic the medium that would normally bathe those cells in vivo. Examples in common use include Eagle's Minimal Essential Medium, Dulbecco's modified Eagle's Minimal Essential Medium (DMEM), Medium 199, RPMI 1640 medium and Ham's F12 Medium. However, virtually no animal cells will grow in such a medium, but require the co-addition of serum. Fetal bovine serum is frequently used as it is more effective than serum obtained from post-natal animals and it contains only minimal concentrations of immunoglobulins which otherwise could have undesirable effects.
The supply of fetal bovine serum is limited by the number of pregnant cows slaughtered. It also has undesirable lot-to-lot variations and may include toxins. Particular concern surrounds its use for the eventual production of recombinant proteins and other pharmaceuticals for human use because the serum may also contain viruses that are harmful to humans and may be carried through a purification protocol that yields the desirable product. Principally for these reasons, extensive efforts have been directed towards the replacement of serum by pure ingredients. Examples of such ingredients are growth factors, hormones and cell attachment factors. Unfortunately, the requirements of each cell type being grown are different and are difficult to establish. Frequently it has not proved possible to achieve equivalent growth properties or equivalent yields of cell products with "serum-free" media as can be obtained with medium containing fetal bovine serum.
The limited availability of fetal bovine serum, its lot-to-lot variability, its resultant considerable cost as well as the deficiencies of "serum-free" media described above have prompted the investigation of other biological fluids as potential replacements in cell culture media. Some progress has been reported in the prior art with bovine milk and bovine colostrum as evidenced by the following selected reports: M. Klagsbrun: "Human milk stimulates DNA synthesis and cell proliferation in cultured fibroblasts" (Proc. Natl. Acad. Sci. USA 75, 5057, 1978); M. Klagsbrun & J. Neumann: "The serum-free growth of Balb/c 3T3 cells in medium supplemented with bovine colostrum" (J. Supramol. Struct. 11, 349, 1979).
The prior art also includes U.S. Pat. No. 4,440,860 to M. Klagsbrun which describes "compositions and methods for promoting cell growth featuring, in one aspect, cell culture media containing milk or colostrum and fibronectin; fibronectin is preferably pre-coated onto the culture substrate" and Japan Patent JP 59166879 to Morinaga "A culture medium for cell incubation--containing milk or milk components". Ultrafiltrates of milk whey have also been used to support the growth of cultured cells, as in European Patent 86401911.2 to G. Linden et al. "Fractions de lait, Procedee d'obtention de ces fractions et milleux de culturo cellulaires renfermant ces fractions" and O. Damerdji et al. "Utilization of whey fractions as a substitute for fetal calf serum in culture media" (Biotech. Tech. 2, 235, 1988).
Despite this progress, a successful alternative to fetal bovine serum is yet to be located.
It is accordingly an object of the present invention to overcome, or at least alleviate one or more of the difficulties or deficiencies related to the prior art.
Accordingly in a first aspect of the present invention there is provided a milk pr

REFERENCES:
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Tissue Growth Factors, ed. by R. Baserga, chapt. 8 pp. 247-276 (1981).
Gaull et al Pediatrics 75(1, part 2) 1985 pp 142-145 Significance of Growth Modulator . . .
Damerdji et al., "Utilization of Whey Fractions as a Substitute for Fetal Calf Serum in Culture Media", Biotech. Tech., 2:235 (1988).
Klagsbrun, "Human Milk Stimulates DNA Synthesis and Cell Proliferation in Cultured Fibroblasts", Proc. Natl. Acad. Sci. USA, 75:5057 (1978).
Klagsbrun et al., "The Serum-Free Growth of Balb/c 3T3 Cells in Medium Supplemented with Bovine Colostrum", J. Supremol. Struct., 11:349 (1979).
Oliver et al., "A Rapid and Convenient Assay for Counting Cells Cultured inMicrowell Plates: Application for Assessment of Growth Factors", J. Cell Biol., 92:513 (1989).
Cox, David A. and Burk, Robert R., Isolation and characterisation of milk growth factor, a transforming-growth factor-.beta.2-related polypeptide, from bovine milk, Eur. J. Biochem. 197:353-358 (1991).
Van Brunt et al., Biotehcnology 6(1):25-30 (Jan. 1988).

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