Chemistry: molecular biology and microbiology – Apparatus
Patent
1993-12-01
1998-01-13
Naff, David M.
Chemistry: molecular biology and microbiology
Apparatus
435176, 435179, 435180, 43524023, 43524024, C12M 100, C12N 500, C12N 1106, C12N 1114
Patent
active
057078599
DESCRIPTION:
BRIEF SUMMARY
BACKGROUND OF THE INVENTION
The present invention relates to a support for the culture of anchorage dependent cells (ADC's) in the shape of microcarriers of a biocompatible material. It relates moreover to a process for preparing such a support.
The mammalian cells used as such for producing human and veterinary vaccines, for obtaining molecules having a high added value and for other analogous production processes, are generally ADC's: they require a support for their growth. Without support, these cells degenerate and die in suspension.
The mass cultivation is realized the better on an industrial scale as the available attachment surface area is larger with respect to the total volume of the used support. A calculation the surface/volume ratio of different supports shows that this ratio can vary between 1.25 and 150.0 for some actual microcarriers (see M. BUTLER, Growth Limitations in Microcarrier Cultures, in: Advances in Biochemical Engineering/Biotechnology, pages 57 and following, volume 34, Springer Verlag, 1997).
On an industrial scale, it is easier to make microcarriers of a spherical geometry: the microbeads. That is the reason why the user has nowadays the choice between a very large variety of this type of microcarriers (see amongst others, A. SHAHAR et al, Nerve and Muscle Cells on Microcarriers in Culture, in: Advances . . . , op. cit., pages 33 and following, volume 34, 1987).
In spite of all their potentialities, there are only a few examples in the world wherein the ADC's are cultivated in mass on microcarriers. The Roux bottles, the rotating flasks and other "cell factories" remain the most generally used systems.
The reasons underlying this situation, as well as the means to be used in order to put an end to this, were the subject of a detailed study (see A. O. A. MILLER et al., Microbeads and Anchorage-Dependent Eukaryotic Cells: The Beginning of a New Era in Biotechnology, in: "Advances . . . op. cit., pages 73 and followings volume 39, 1989).
Recently, essentially in order to reduce the production costs of substances produced by the ADC's, use has been made of polystyrene spirals and of disks having a thickness of 1.5 mm and a diameter of 6 mm formed by a polypropylene netting coated with a polyester film (see A. KADOURI et al., Polystyrene substratum for bulk culture of anchorage dependent cells, in Cytotechnology, 1: 301-307, 1988; M. NEEMAN et at., Adaptation of Culture Methods for NMR Studies of Anchorage-Dependent Cells, in Magnetic Resonance in Medicine, 7, 236-242, 1988; A. KADOURI et al., Production of Anti-Leukemic Factor from Stroma Cells in a Stationary Bed Reactor on a New Cell Support, in 9th ESACT Meeting, Belgium, pages 327 and following, Butterworths, 1989).
All the hereabove mentioned supports are characterized by a three-dimensional geometry. For many of these supports, even with respect to the microbeads and the thick disks of a woven material, the surface/volume ratio of the support remains unfavourable. Besides, these thick disks made of a netting of a woven material present the drawback of allowing the cells to penetrate and to attach within these disks. These cells can no longer be removed subsequently from their support or with very much difficulties, which represents an important drawback and a loss of dry matter when the cells are cultivated for themselves. As a matter of fact, these disks allow only a culture of cells attached ad vitam to their support, for example for the purpose of secreting a given substance which is collected. In this type of culture, the cell supports are not in suspension.
BRIEF DESCRIPTION OF THE DRAWINGS
FIGS. 1 to 3 represents microscopic photo of three reference disks, respectively, made of a cellophane quality A film, a cellophane quality B and a polystrene film, and subjected to the Tests later described in this application.
FIGS. 4 to 6 represents similar photos of three disks made of the same films as in FIGS. 1 to 3 but after projection in a magnetron of tinanium plasma onto both their surfaces.
FIGS. 7 to 9 represents sim
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A. Johansson and V. Nielsen (1979) Develop. biol. Standard, Biosilon R A New Microcarrier, 46, pp. 125-129 (S. Karger, Basel 1980).
De Hollain Guy
Masquelier Dominique
Miller Alain O. A.
Naff David M.
Nunc A/S
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