Chemistry: molecular biology and microbiology – Apparatus – Bioreactor
Reexamination Certificate
2001-06-15
2003-05-27
Beisner, William H. (Department: 1744)
Chemistry: molecular biology and microbiology
Apparatus
Bioreactor
C435S395000, C435S304300, C435S294100
Reexamination Certificate
active
06569675
ABSTRACT:
BACKGROUND OF THE INVENTION
1. Field of the Invention
The present invention relates in general to the cellular biological field and, in particular, to a cell cultivating flask and method for using the cell cultivating flask to grow cells.
2. Description of Related Art
Manufacturers of cell cultivating flasks have been for some time trying to design a cell cultivating flask that is more simple and convenient to use than the traditional cell cultivating devices. A cell cultivating flask is basically a device which contains one or more trays that are designed to receive a cell cultivating media and to enable the growth of cells within the cell cultivating media. Examples of two traditional cell cultivating devices are briefly discussed below and described in U.S. Pat. Nos. 5,310,676 and 4,172,013 both of which are hereby incorporated by reference herein.
Referring to
FIGS. 1A and 1B
(PRIOR ART), there are illustrated two perspective views of a traditional cell cultivating device described in the aforementioned U.S. Pat. No. 5,310,676. The cell cultivating device shown in
FIGS. 1A and 1B
comprises an outer container or flask
10
, which is made from a transparent or non-transparent material, such as glass or plastics material, or from metal. The container
10
is provided with a neck
11
defining a filling opening
12
. The neck
11
is formed with outer screw threads
13
for cooperating with inner screw threads of a screw cap
14
by means of which the filling opening
12
may be closed. The outer container
10
has a flat bottom wall
18
, a top wall
19
, opposite side walls
20
, a flat end wall
21
, and an opposite end wall
22
on which the neck
11
is formed.
A number of partition wall members
15
having a bottom wall or cell attachment plate
16
and a surrounding peripheral side wall
17
extending upwardly therefrom, is arranged in superposed relationship within the container
10
, so as to define cultivating chambers therein. The lower partition wall member
15
arranged adjacent to the bottom wall
18
is supported by feet or spacer members
23
so that a cultivating chamber is defined between the inner surface of the container bottom wall
18
and the lower surface of the bottom wall
16
of the lower partition wall member
15
. The spacer members
23
may be formed as integral parts of the walls of the outer container
10
or of the lower partition wall member. An upper cultivating chamber is defined between the inner surface of the container top wall
19
and the upper surface of the adjacent bottom wall
16
of the upper partition wall member
15
. Furthermore, a cultivating chamber is defined between each pair of adjacent partition wall members
15
. Thus, the cell cultivating device shown in
FIGS. 1A and 1B
which comprises two partition wall members
15
defines three cultivating chambers therein.
The container end wall
22
has a convex contour so that a manifold chamber
24
is defined between the container end wall
22
and the adjacent side walls
17
of the partition wall members
15
. Cut-outs in the peripheral side walls
17
of the partition wall members
15
define overflow openings
26
communicating with the manifold chamber
24
.
The outer container
10
may be made from at least two separate container parts, which may, for example, be molded from transparent plastics material. When the partition wall members
15
have been arranged within one of the container parts, such container parts may be sealingly interconnected, for example by interconnecting flanged rim portions of the container parts so as to form a heat seal
27
in a plane being substantially parallel with the bottom walls
16
of the partition wall members
15
.
As indicated in broken lines in
FIGS. 1A and 1B
each of the partition wall members
15
further comprises an equalizing opening
30
formed in its peripheral side wall
17
adjacent to the container end wall
21
. The equalizing opening
30
may be a cut-out formed in the rim portion of a curved part of the peripheral wall
17
of each partition wall member. The curved parts of the superposed partition wall members
15
define a transverse tunnel or passage
31
interconnecting the superposed cell cultivating chambers. The tunnel or passage
31
may have any suitable cross-sectional shape, which may, for example, be a semi-circle as shown.
When the cell cultivating device shown in
FIGS. 1A and 1B
is to be used it is positioned in its upright position shown in
FIG. 1A
, and the screw closure cap
14
is removed. A suitable amount of cell cultivating medium and cells to be cultivated are poured into the manifold chamber
24
of the container
10
through the filling opening
12
defined by the neck
11
. From the manifold chamber
24
of the liquid cell cultivating medium flows into the various cultivating chambers via the overflow opening
26
and through the opening defined between the inner surface of the container bottom wall
18
and the bottom wall
16
of the adjacent partition wall member
15
. Now, the level of the cell cultivating media within the cultivating chambers will be the same after a short period of time because the cultivating chambers are all interconnected by the tunnel
31
and the openings
30
formed therein. Provided that the dimensions of the cultivating chambers are substantially the same, these chambers will now contain substantially the same amount of cell cultivating media.
After the filling opening
12
has been closed by the screw cap
14
, the flask
10
may then by a quick movement be tilted from the upright position shown in
FIG. 1A
to the position shown in
FIG. 1B
in which the container bottom wall
18
is supported in a substantially horizontal position. As the equalizing openings
30
are relatively small, this simple procedure renders it possible to have substantially the same amount of cell cultivating medium placed in all of the now horizontally extending cultivating chambers. This means that a layer of cell cultivating medium containing cells to be cultivated is supported by the container bottom wall
18
and by the bottom walls or cell attachment plates
16
of each of the partition wall members
15
. The surfaces of these walls have preferably been subject to a surface treatment allowing good cell attachment.
After expiration of the cultivating period the cell cultivating medium may be poured out through the inlet opening
12
of the neck
11
. Thereafter, the cells attached to the bottom walls
16
and
18
may be scraped or flushed out through the inlet or filling opening
12
. Alternatively, the container bottom wall
18
or top wall
19
, or any of the container side or end walls
20
and
21
, respectively, may be cut away or otherwise removed so that the tray members may be taken out from the container
10
, whereafter the cultivated cells may be scraped or flushed from the bottom walls
16
and
18
.
A main drawback of this traditional cell cultivating device is that a user does not have direct access to each of the cultivating trays. In other words, the user can not fit a pipette of any size onto any cultivating tray to add or remove the cell cultivating media. Of course, it would be desirable to enable the user to have direct access to each of the cultivating trays. Another drawback of this traditional cell cultivating device is that the cell cultivating media often wicks via capillary action between the inner walls of the outer container
10
and the outer walls of the wall members
15
. Cells that are trapped in this manner can easily die and contaminate the other cells.
Referring to
FIGS. 2A and 2B
(PRIOR ART), there are illustrated two sectional front views of a traditional cell culture system described in the aforementioned U.S. Pat. No. 4,172,013. The traditional cell culture system can be used to enable the mass growth of cell by introducing a nutrient medium, cell suspension material etc. through a central feed line
1
serving as supply channel into the lower part of a system of communicating chambers
2
. As shown in
FIG. 2A
, the system is in a
Lyman George F.
Wall Joseph C.
Beisner William H.
Corning Incorporated
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