Chemistry: molecular biology and microbiology – Apparatus – Mutation or genetic engineering apparatus
Patent
1990-08-13
1991-11-12
Lacey, David L.
Chemistry: molecular biology and microbiology
Apparatus
Mutation or genetic engineering apparatus
435310, 435240242, 435813, 21050023, 21050025, 2103218, C12M 300, B01D 6300, B01D 3600
Patent
active
050647644
DESCRIPTION:
BRIEF SUMMARY
DESCRIPTION
The present invention relates to a bioreactor and to an apparatus for the continuous culture on an industrial scale of animal cells. It is applicable to the production of all animal cells and to the production of metabolites produced by the cells, such as e.g. monoclonal antibodies. The bioreactor and the apparatus according to the invention permit the culture under sterile conditions of animal cells, such as hybridomas.
Various bioreactors are already known, which can be used in the culture of free cells (non-adhering), like those described in EP-A-0 113 328, EP-A-0 112 155, U.S. Pat. No. 4,661,455, EP-A-0 155 237 and EP-A-0 112 154. In all these known bioreactors use is made of water-impermeable, but gas-permeable membranes, so as to ensure the necessary oxygen transfer for the development of any animal cell. During the size extrapolation of these reactors, the problem of transfer surfaces very rapidly occurs. For example, it is calculated that for a Braun-type reactor equipped with a stirred vessel and silicone, oxygen transfer tubes, 700 meters of tubing would be required for a 75 l fermenter. In addition, the last four documents referred to hereinbefore relate to reactors constituted by planar membranes suffering from the disadvantage of bringing about non-homogeneous transfers.
Moreover, there is no way in which it is possible to regulate and control the clogging of these membranes by the cultured cells. In particular, the first aforementioned document forces the passage of the solution containing the cells through membranes. The immediate result is a much larger transfer of substances, but a side effect is an increase in the transfer pressure and a compression of the polarizing layer which has formed on the membranes and this has the consequence of increasing the risk of said membranes becoming blocked.
Moreover, apparatuses based on the confinement of cells between two different membrane types, one used for supplying certain substrates to the cells and the others for extracting certain products, have no device for regulating and checking on an intermediate basis the pressure drop and consequently have membrane portions which are poorly used, the pressures being close to one another on either side of the said membranes in certain areas, which leads to a limited transfer, which does not permit a controlled, intense supply of the necessary nutrient substrates.
Moreover, the progressive clogging of the membranes leads to a simultaneous reduction in the transfers of all the nutrient substrates, but in often varying proportions, which is prejudicial to the checking of the concentrations over a period of time and makes it impossible for these reactors to operate in a satisfactory manner over long periods.
The invention specifically relates to a bioreactor and to an apparatus for the culture of animal cells making it possible to obviate the disadvantages referred to hereinbefore.
The present invention therefore relates to a bioreactor for the culture of animal cells comprising:
a cellular culture chamber, formed by a sleeve and facing walls of two enclosures, respectively the intake enclosure and the discharge enclosure, containing on each occasion the cells and the culture medium, each enclosure being provided with an internal partition subdividing said enclosure into a first and a second respectively internal and external chambers;
at least one first porous mineral tube connecting the external chamber of the intake enclosure to the internal chamber of the discharge enclosure and traversing said culture chamber for supplying nutrient medium to the culture medium, said first porous mineral tube having on its inner face a first microporous filtering membrane allowing the passage of molecules of the nutrient medium with an average or middle molecular weight, but stopping the macromolecules of a high molecular weight, and on its outer face a second microfiltering membrane permitting the passage of the low molecular weight molecules and serving as a barrier for the cells and most of the proteins p
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patent: 4722902 (1988-02-01), Harm et al.
patent: 4804628 (1989-02-01), Cracauer et al.
patent: 4849104 (1989-07-01), Garcera et al.
patent: 4874516 (1989-10-01), Kondo
patent: 4938931 (1990-07-01), Cussler
Besnainon Bernard
Elluard Marie-Paule
Lessart Pierre
Philippe Alain
Chan William K. Y.
Commissariat a l''Energie Atomique
Lacey David L.
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