Chemistry: molecular biology and microbiology – Carrier-bound or immobilized enzyme or microbial cell;... – Enzyme or microbial cell is immobilized on or in an organic...
Patent
1996-03-06
1999-09-07
Naff, David M.
Chemistry: molecular biology and microbiology
Carrier-bound or immobilized enzyme or microbial cell;...
Enzyme or microbial cell is immobilized on or in an organic...
435174, 435180, 435182, 435395, 435400, 4352891, 4352971, 4352974, C12N 1102, C12N 500, C12N 506, C12M 300
Patent
active
059486554
DESCRIPTION:
BRIEF SUMMARY
FIELD OF THE INVENTION
The invention relates to a process for treating cell cultures, in particular hepatocytes on hollow fibres, which are at least partially gas and/or liquid permeable, whereby the cell cultures are applied on or into the hollow fibres and suspension medium is conveyed as a culture medium through or around the hollow fibres. The invention also relates to a device for performing the process.
BACKGROUND OF THE INVENTION
In medicine and in pharmacognosy it is frequently necessary to perform experiments with cell cultures, in particular with liver cells (hepatocytes). This applies, for example, to their culture, their observation, in particular the observation of reactions on foreign and/or poisonous substances, to storage and the like.
Furthermore the quest for suitable organ replacement is becoming increasingly important.
In known systems for the culture of liver cells in so-called hollow fibre reactors, the cells were previously introduced in liquid form either inside or outside the hollow fibres by suspension in a solution, such as, e.g. a culture medium. After the adhesion of these cells to the fibres or also by leaving non-adherent cells in the interspaces between the adhered cells in the form of an aggregate formation, the system was used for the respective purpose.
The crucial disadvantage of the known processes is that intercellular contacts between the individual hepatocytes are only developed in an inadequate manner or in some cases not at all.
The loss of cellular morphology not only entails a loss of function, but also prevents regenerative power after a stimulation of the cells with growth factors in a longer-term culture system. Longer-term cultivation would however be of great significance with the use of human hepatocytes, as they are not available in any quantity.
SUMMARY OF THE INVENTION
The object of the present invention is to create a process of the type mentioned at the beginning with which a mass culture is possible, whereby one wishes to achieve, as far as possible, an "in vivo" state, in particular the longest possible life-span.
This object is achieved in accordance with the invention in that the cells of the cell cultures are covered with an outer matrix layer on the side faced away from the hollow fibres after their application and adhesion to the hollow fibres, directly or indirectly over a first matrix layer.
With the process in accordance with the invention, the cell cultures are embedded in an extra-cellular matrix layer in the manner of a sandwich technique known per se. At the same time a reorganisation of the cell shape and a renewed formation of microvilli on the side or sides faced towards the matrix surfaces occur. This corresponds to the natural shape of the liver cells and promotes exchange of substances in the manner of sinusoidal membranes. However, apart from these morphological advantages, the functional preservation of these cells is also important.
One of the most important features of the invention is consequently represented by the second, i.e. the outer, matrix layer, which envelops the cell cultures. This enveloping is however only performed when the cell cultures are applied to the hollow fibres and have adhered there. This can occur directly or over an intermediate first matrix layer. The use of a first matrix layer depends on what type of cell cultures are to be treated and what material is used for the hollow fibres. In connection with hollow fibres, specific cell cultures require no separate matrix layers as an intermediate layer, but can adhere directly to the surfaces of specific hollow fibres.
In an advantageous embodiment of the invention it may be specified that the cells are applied to a plurality of hollow fibres on their outer side in each case, with the cells being covered on the outside by the matrix layer in each case.
Depending on the diameter of the hollow fibres, the cell cultures and the matrix layer or respectively the matrix layers may also be synthesised internally in the cavities of the hollow fibres. However f
REFERENCES:
patent: 5605835 (1997-02-01), Hu et al.
patent: 5658797 (1997-08-01), Bader
Patent Abstracts of Japan-13 vol. 14, No. 254 (C-0724), May 31, 1990.
Nyberg et al., Bilirubin Conjucation in a Three Compartment Hollow Fiber Bioreactor, Proceedings of the 12th Annual International Conference of the IEEE Engineering in Medicine and Biology Society, vol. 12: Part 1/5, Pederson et al., Eds., pp. 443-444, Nov. 1-4, 1990.
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