CELL CULTURE BOTTLE

Abstract

The invention relates to a cell culture bottle for adherent cells (e.g. human mesenchymal stem cells), comprising: a vessel; an internal cylinder, which has an internal Archimedes screw; an internal central tube, through which liquid can flow; and at least one wall arranged around the central tube. This arrangement provides an enlarged inner surface for the growth of the cells and for the reliable mixing of the fluid. The cell culture bottle is formed as a single piece and can be simply and economically produced as a disposable device by means of additive manufacturing.

Claims

1. A cell-culture bottle comprising a vessel comprising three subregions: a base region, a lid region and a cylinder region arranged therebetween, an Archimedes' screw arranged internally in the cylinder region of the vessel and having a central tube which is arranged along the central axis of said Archimedes' screw and through which liquid is flowable, at least one wall which is arranged concentrically around the central tube and which extends over the entire cylinder region of the vessel, wherein the cell-culture bottle is formed as one piece and wherein the central tube is designed such that it has a first opening on the outer side of the vessel in the lid region and has a second opening inside the vessel on a bottom side of the cylinder region and wherein the Archimedes' screw is integrally connected to the cylinder region of the vessel and to the at least one wall, wherein the Archimedes' screw and the at least one wall are arranged such that a fluid can pass along the Archimedes' screw from the bottom side of the cylinder region into the lid region of the vessel upon rotation of the cell-culture bottle.

2. The cell-culture bottle as claimed in claim 1, wherein the vessel has an opening in the lid region beyond the central tube.

3. The cell-culture bottle as claimed in claim 1, wherein the cell-culture bottle has a coating comprising polysaccharides and/or peptides, wherein said coating is applied on a surface of the Archimedes' screw, the at least one wall and/or an internal side of the vessel, so that cells can adhere to the inner surfaces of the cell-culture bottle and grow thereon.

4. The cell-culture bottle as claimed in claim 1, it further comprising a closed base region, meaning that the cell-culture bottle is designed to be closed on the bottom side of the cylinder region, wherein the base region is integrally connected to the cylinder region.

5. The cell-culture bottle as claimed in claim 1, wherein the cell-culture bottle has a rotation mechanism for rotation of the vessel, wherein said rotation mechanism is attachable to the base region of the vessel.

6. The cell-culture bottle as claimed in claim 1, wherein the cell culture bottle has a filter in the central tube.

7. The cell-culture bottle as claimed in claim 1, wherein the central tube has a third opening, wherein said third opening is located at a point of contact between the central tube and the lid region of the vessel, so that fluid can get from the base region to the lid region of the vessel via the Archimedes' screw and can, via the third opening into the central tube 40, flow back to the bottom side of the cylinder region of the vessel.

8. The cell-culture bottle as claimed in claim 1, wherein the cell-culture bottle it is made of a material suitable for additive manufacturing.

9. The cell-culture bottle as claimed in claim 8, wherein the material is selected from the group consisting of acrylonitrile butadiene styrene (ABS), acrylate styrene acrylonitrile (ASA), high-impact polystyrene (HIPS), thermoplastic elastomers (TPE), polycarbonates (PC), polylactides (PLA), polyethylene terephthalate (PET), polyethylene terephthalate glycol (PETG), polymethyl methacrylate (PMMA) or polyvinyl alcohol (PVA or PVOH).

10. A culturing system, comprising a cell-culture bottle as claimed in claim 1.

11. The use of a cell-culture bottle as claimed in claim 1 for culturing of cells.

12. A method for culturing cells in a cell-culture bottle as claimed in claim 1, comprising at least the steps of: a) filling the cell-culture bottle with fluid; b) adding the cells to be cultured to the cell-culture bottle; c) rotating the cell-culture bottle, so that the fluid is distributed and covers the cells, so that the cells can settle in the vessel, in the Archimedes' screw, on the at least one wall and the central tube and divide; d) removing the fluid at a certain cell density; e) adding a buffer to the cell-culture bottle in order to wash the cells; f) detaching the cells from the vessel, the Archimedes' screw, the at least one wall and the central tube; g) stopping the detachment reaction by addition of fresh fluid; and h) removing the fluid containing detached cultured cells.

Description

FIGURE LEGENDS AND LIST OF REFERENCE SIGNS

[0065] FIG. 1 shows, by way of example, one embodiment of the cell-culture bottle 1 according to the invention. Here, the closed base region 12 shown is optional. Furthermore, the third opening 43 of the central tube 40 is optional.

[0066] FIG. 2 shows an exemplary arrangement of multiple walls 50 around the central tube 40.

[0067] The remaining components of the cell-culture bottle 1 are not shown in this figure for the sake of clarity.

[0068] FIG. 3 shows, by way of example, in a side view, FIG. 3a, an Archimedes' screw 30 having a central tube 40. FIG. 3b displays the longitudinal section.

[0069] FIG. 4 shows how much the growth surface area F in the vessel 10 increases in relation to the radius R of the vessel 10. In this example, the cylinder region 16 has a height of 16 cm to 20 cm and comprises 20 turns, with a distance of 0.8 cm between the walls 50.

[0070] FIG. 5 shows the use of the cell-culture bottle 1 according to the invention instead of a stirrer in an inclined classic bioreactor.

LIST OF REFERENCE SIGNS

[0071] 1 cell-culture bottle [0072] 2 reaction tank [0073] 3 probes [0074] 4 inflow [0075] 5 outflow [0076] 6 fluid [0077] 7 rotation mechanism of the stirred tank reactor [0078] 10 vessel [0079] 11 opening of the vessel 10 [0080] 12 base region [0081] 14 lid region [0082] 16 cylinder region [0083] 30 Archimedes' screw [0084] 40 central tube [0085] 41 first opening of the central tube 40 [0086] 42 second opening of the central tube 40 [0087] 43 third opening of the central tube 40 [0088] 50 wall [0089] α angle of inclination of the Archimedes' screw 30 [0090] β angle of inclination of the vessel 10 [0091] D diameter of the vessel 10