Chemistry: molecular biology and microbiology – Apparatus – Differentiated tissue perfusion or preservation apparatus
Patent
1990-02-05
1991-09-03
Lacey, David L.
Chemistry: molecular biology and microbiology
Apparatus
Differentiated tissue perfusion or preservation apparatus
435813, 435819, 435286, 43524025, 43524046, 422224, C12M 300, C12N 500
Patent
active
050454706
DESCRIPTION:
BRIEF SUMMARY
CROSS REFERENCE TO RELATED APPLICATIONS
This application is a national phase application of PCT/CH89/00067 filed Apr. 4, 1989 and based upon a Swiss National Application 1244/88-6 filed Apr. 5, 1988 under the International Convention.
FIELD OF THE INVENTION
The invention relates to a device for submerged culture of vegetable, animal and human tissue cells within a closed chamber, consisting of a container that can be sterilized and is equipped with a centrally disposed, rotatable hollow shaft having one or more stirring elements and baffles arranged one above the other.
BACKGROUND OF THE INVENTION
For the submerged culture of tissue cells the standard stirrers and mixers used in the cultivation of microorganisms have proved to be unsuitable due to the shearing forces which were generated during their operation. Particularly when cultivating human cell lines, the strong mechanical movements lead to inadmissible damage to the cells. One has therefore carried out the exchange of nutrients and gases by the so-called airlift method whereby the aerating medium took care of the necessary mixing by means of passing through perforated plates. Also a combination of pneumatic and mechanical mixing was tried.
In particular, EP-A-0 095 804 describes a method for the submerged culture of tissue cells whereby a stirrer operating at a relatively low speed provides for a thorough mixing of nutrients and air, while a sintered plate arranged on the bottom of the container provides of the gassing. This known device is disadvantageous because the gas transfer to and from the cell is insufficent since gassing within a restricted space can take place only from the bottom of the container. The relatively slow motion of the stirrer prevents a sufficient aeration in the area of the container walls and in the upper zone of the container.
OBJECT OF THE INVENTION
It is an object of the invention to provide a device that permits an optimal gassing in a cultivation container and avoids damage to the cells.
SUMMARY OF THE INVENTION
The device, according to the invention, has characterized in baffle plates arranged between each pair of stirring elements and equipped with a gas-impermeable part and a gas-permeable part.
This device has the advantage that the gassing can be effected with high intensity on different levels up to the surface of the liquid, in spite of the slow-moving stirrer. The element-type construction of the stirring elements on the hollow shaft permits their assembly in containers of different dimensions, depending upon the kind of cells to be cultivated. Across the hollow stirring elements an optimal gassing is thus assured on each level without the danger of cells being damaged due to shear forces generated by a high-speed stirrer.
It is particularly advantageous to make the upper part of the stirring elements of high-grade stainless steel sheet, which has proved to be particularly suitable. The gas apertures may be round or of any other suitable shape.
It is advantageous to provide the upper part of the stirring elements as a porous body whereas the lower part is closed. On the other hand the entire stirring element may consist of a porous material.
As a porous material sintered metal, ceramics and synthetic material are suitable. It synthetic material is used, it must be thermially stable enough so as to withstand several sterilizations at temperatures of about 130 centigrades without becoming damaged.
For particular cell cultivations, it has proved to be suitable to provide the gas-permeable part of the baffle plates with a semipermeable diaphragm. Such a diaphragm permits, besides the gas transfer, also the supply of nutrient medium.
In addition it has proved to be advantageous to arrange the baffle plates between a pair of stirring element within a removable baffle plate cage. The baffle plate cage may be cleaned separately and pre-sterilized if required. In this manner slow-moving stirrers having a great surface area and provided with gas passages on their upper side may rotate within a baffle plate
REFERENCES:
patent: 2121458 (1938-06-01), Vogelbusch
patent: 4379846 (1983-04-01), Shkidchenko et al.
patent: 4494878 (1985-01-01), Rainey, Jr.
Composite Sciences Inc. in Chemical Eng., 81(6), p. 80.
Chan William K. Y.
Dubno Herbert
Lacey David L.
Stawag
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