Oligopeptide-free cell culture media

Inactive Publication Date: 2007-09-13
BAXTER HEALTHCARE SA +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0013] It is an object of the present invention to provide oligopeptide-free cell culture media. A further object of the present invention is to provide methods for cultivating cells in said media as well as methods for efficient expression of recombinant proteins and/or methods for the efficient production of viruses.
[0014] It is another object of the present invention to eliminate animal, plant and/or yeast derived hydrolysates and to provide media which do not comprise any added supplementary proteins or oligopeptides.
[0015] Surprisingly, the addition of at least 0.5 mg/L of a polyamine to cell culture media provides an advantageous effect not on

Problems solved by technology

Moreover, bovine serum and products derived thereof bear the risk of BSE contamination.
In addition, all serum-derived products can be contaminated by unknown substances.
When using serum or protein additives derived from human or animal sources in cell culture, there are numerous problems (e.g., the varying quality in composition of different batches and the risk of contam

Method used

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  • Oligopeptide-free cell culture media
  • Oligopeptide-free cell culture media
  • Oligopeptide-free cell culture media

Examples

Experimental program
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Effect test

example 1

Preparation of BAV-Medium

[0072] Oligopeptide-free medium (BAV-medium) was prepared with basal DMEM / HAM's F12 (1:1) comprising 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: L-Asparagine.H2O 20 mg / L, L-Cysteine.HCl.H2O 15 mg / L, L-Cystine.2 HCl 20 mg / L, L-Proline 35 mg / L, L-Tryptophan 20 mg / L.

example 2

Determination of Cell Counts

[0073] 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 fluorescent 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

example 3

Determination of FVIII Activity

[0074] The activity of Factor VIII (FVIII) (cf. FIGS. 1 to 5) was measured by a chromogenic assay (Chromogenic, Sweden).

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Abstract

The present invention relates to oligopeptide-free cell culture media comprising at least 0.5 mg/L of a polyamine and to methods for cultivating cells in said oligopeptide-free cell culture media comprising at least 0.5 mg/L of a polyamine. The invention also relates to methods for expressing at least one protein in a medium comprising at least 0.5 mg/L of a polyamine and to methods for producing at least one virus in a medium comprising at least 0.5 mg/L of a polyamine.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS [0001] This application claims benefit of U.S. provisional application No. 60 / 756,419, filed Jan. 4, 2006, which application is herein incorporated by reference.FIELD OF THE INVENTION [0002] The present invention relates to oligopeptide-free cell culture media comprising at least 0.5 mg / L of a polyamine and to methods for cultivating cells in said oligopeptide-free cell culture media comprising at least 0.5 mg / L of a polyamine. The invention also relates to methods for expressing at least one protein in a medium comprising at least 0.5 mg / L of a polyamine and to methods for producing at least one virus in a medium comprising at least 0.5 mg / L of a polyamine. BACKGROUND OF THE INVENTION [0003] For cultivation of cells, particularly eukaryotic cells, and more specifically mammalian cells, there is a constant need to use special culture media providing nutrient substances that are required for efficient growth of cells and for the production of b...

Claims

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Application Information

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IPC IPC(8): C12N7/00C12N1/20C12N7/01C12N5/00C12N5/07C12N5/071C12N5/077
CPCC12N5/0031C12N7/00C12N2500/05C12N2500/10C12N2710/24151C12N2500/32C12N2500/38C12N2500/46C12N2500/50C12N2500/24C12N5/005C12N5/00C12N5/0018C12N2500/20C07K14/755
Inventor GRILLBERGER, LEOPOLDREITER, MANFREDMUNDT, WOLFGANGMITTERER, ARTUR
Owner BAXTER HEALTHCARE SA
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