Animal protein-free media for cultivation of cells

a technology of protein-free cell culture and animal protein, which is applied in the field of animal protein-free cell culture media, can solve the problems of bse contamination of all serum-derived products, risk of contamination with mycoplasma, and contaminated products derived from them, so as to increase the protein expression per cell, promote cell growth, and increase the yield of desired products

a technology of protein-free cell culture and animal protein, which is applied in the field of animal protein-free cell culture media, can solve the problems of bse contamination of all serum-derived products, risk of contamination with mycoplasma, and contaminated products derived from them, so as to increase the protein expression per cell, promote cell growth, and increase the yield of desired products

US20060094104A1Inactive Publication Date: 2006-05-04BAXTER INT INC +1
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  • Animal protein-free media for cultivation of cells
  • Animal protein-free media for cultivation of cells
  • Animal protein-free media for cultivation of cells

Examples

Experimental program
Comparison scheme
Effect test

example 1

BAV-Medium

[0071] Animal protein free medium was prepared with basal DMEM / HAM's F12 (1:1) medium supplemented with inorganic salts, amino acids, vitamins and other components (Life technologies, 32500 Powder). Also added were L-glutamine (600 mg / L), ascorbic acid (20 μM), ethanol amine (25 μM), Synperonic® (SERVA) (0.25 g / L), sodium selenite (50 nM). Additionally, essential amino acids were supplemented to the cell culture medium. Further, varying concentrations of soy hydrolysate (Quest Technologies, NY or DMV Intl., NY ) in the range of 0.0-1.0% and varying concentrations of polyamines (0-10 mg / L) were added (FIG. 1-9)

example 2

[0072] Cell cultures of recombinant mammalian cells (e.g. CHO-cells stably expressing Factor VIII=GD8 / 6-cells) were grown in suspension in a chemostat culture in 10 I bioreactors. The culture conditions of 37° C., oxygen saturation 20% and pH 7.0 to 7.1 were kept constant. The cultures were supplied with a constant feed of BAV-medium as defined in Example 1 additionally supplemented with soy hydrolysates in the range of 0.1-1.0% and / or addition of putrescine.2HCl in the range of 0-1 mg / L (cf. FIG. 1-5).

[0073] Small scale experiments with GD8 / 6 cells in suspension culture were carried in Techne spinner flasks at 200 ml working volume in batch refeed mode at 37° C., without pH and pO2 control. The cultures were supplied with BAV-medium as defined in Example 1 without supplementation of soy hydrolysate and polyamines, or supplemented with soy hydrolysate in the range of 0.1-0.4% and / or putrescine.2HCl, ornithine.HCl, spermine.4HCl in the range of 0-18 mg / L (equivalent to 0-10 mg / L of ...

example 3

cf. FIG. 1 to 57, and 9

[0074] Cell counts from suspension cells or immobilized cells were determined either by counting with a CASY® cell counter as described by Schärfe et al., (Biotechnologie in LaborPraxis 10: 1096-1103 (1988)) or by citric acid extraction and flourescent staining of the nuclei followed by counting with a NucleoCounter® (Chemometec, DK). The specific growth rate (μ) is calculated from the increase of the cell densities (Xt) and / or the dilution rate (D) of the steady state of chemostat cultures of suspensions cells over a certain time interval (t):

μ=D+In(Xt / X0) / t

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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.

Description

FIELD OF THE INVENTION [0001] The present invention relates to animal protein-free cell culture media comprising a polyamine 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. BACKGROUND OF THE INVENTION [0002] For cultivation of cells, particularly eukaryotic cells, and more specifically mammalian cells, there is a constant need to use special culture media that make available the growth nutrient substances that are required for efficient growth of the cells and for the production of the proteins or viruses that are desired. For the efficient production of biological products, such as viruses or recombinant proteins, it is i...

Claims

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Application Information

Patent Timeline
04 May 2006
Publication
US20060094104A1
IPC
C12N7/00; C12N5/02
CPC
C12N5/0031; C12N2500/46; C12N2500/74; C12N2500/76; C12N2500/99; C12N5/0043; C12N2500/92; C12N5/00
Inventors
GRILLBERGER, LEOPOLD; REITER, MANFRED