FLEXIBLE BAG FOR CULTIVATION OF CELLS

• US 20140011270A1
• Filed: 03/16/2012
• Published: 01/09/2014
• Est. Priority Date: 03/18/2011
• Status: Active Grant

• Current Assignee: Cytiva Sweden AB (Danaher Corporation)
• Original Assignee: GE Healthcare Bio-Sciences AB (General Electric Company)
• Inventors: Kaisermayer, Christian, Lindskog, Eva, Walsh, Kieron D., Chotteau, Veronique, Diana, Rafael, Robinson, Craig, Rucker, Jimmie L.
• Granted Patent: US 9,550,969 B2
• Time in Patent Office: Days
• US Class Current: 435/326
• CPC Class Codes: B01F 31/23 by pivoting the containers about an axis
• CPC Class Codes: B01F 35/513 Flexible receptacles, e.g. bags supported by rigid containers
• CPC Class Codes: C12M 23/14 Bags
• CPC Class Codes: C12M 27/16 Vibrating; Shaking; Tilting
• CPC Class Codes: C12M 27/20 Baffles; Ribs; Ribbons; Auger vanes

Abstract

An inflatable bioreactor bag for cell cultivation, which comprising a top and a bottom sheet of flexible material, joined together to form two end edges and two side edges, wherein one baffle or a plurality of baffles extend from the bottom sheet in a region where the shortest distance to any one of the two end edges is higher than about one fourth of the shortest distance between the two end edges.

Claims

1. An inflatable bioreactor bag (1) for cell cultivation comprising a top (2;

22) and a bottom (3;

23) sheet of flexible material, joined together to form two end edges (4) and two side edges (5), wherein one baffle (6) or a plurality of baffles (6) extend from said bottom sheet in a region where the shortest distance to any one of said two end edges (4) is higher than about one fourth of the shortest distance between said two end edges.

2. The inflatable bioreactor bag of claim 1, wherein said baffles (6) are joined to said bottom sheet (3;

23) and to said top sheet (2;

22).

3. The inflatable bioreactor bag of claim 1, wherein said baffles (6) are generally cylindrical, such as with circular or elliptic cross sections.

4. The inflatable bioreactor bag of claim 1, wherein said bag is equipped with sterile connectors and irradiated.

5. The inflatable bioreactor bag of claim 1, wherein at least one, such as each, baffle has a generally sigmoidal shape.

6. The inflatable bioreactor bag of claim 1, wherein at least one of said baffles (6), such as two baffles, is/are tubular.

7. The inflatable bioreactor bag of claim 6, wherein said tubular baffle(s) is/are joined to said bottom sheet via one cell suspension diffuser per baffle and to said top sheet via one inlet/outlet port per baffle.

8. The inflatable bioreactor bag of claim 7, wherein at least one, such as each, cell suspension diffuser (10;

20;

30) comprises a bottom ring (24;

34) or plate, a plurality of struts (25;

35) joining said bottom ring or plate to a tube connector (27;

37) and a plurality of openings (26;

36) between said struts.

9. The inflatable bioreactor bag (41;

51) of claim 7, wherein at least one of said ports (42;

52) on said top sheet is further connected to at least one cell separating device, such as a filter (43;

53) or a gravitational device, outside the bag (41;

51).

10. The inflatable bioreactor bag (41;

51) of claim 9, wherein said filter (43;

53) is a microfilter or an ultrafilter with a retentate side and a permeate side, such as a hollow fiber cartridge.

11. The inflatable bioreactor bag of claim 9, wherein said cell separating device is further connected to a reciprocating pump (44), such as an air actuated membrane pump.

12. The inflatable bioreactor bag of claim 10, wherein two of said ports (52) on said top sheet are connected via a pump (54) to the retentate side of said filter (53), forming a retentate loop.

13. A bioreactor comprising the inflatable bioreactor bag (1) of claim 1, mounted on a support (8) which is pivotally mounted to a base (9) about a movable axis (7) generally parallel to said end edges (4).

14. A method for cell cultivation comprising the steps of:

a) providing an inflatable bioreactor bag comprised of a top and a bottom sheet of polymer material, joined together to form two end edges (4) and two side edges (5), wherein one baffle (6) or a plurality of baffles (6) extend from said bottom sheet in a region where the shortest distance to any one of said two end edges is higher than about one fourth of the distance between said two end edges;

b) mounting said bioreactor bag on a support pivotally mounted to a base about a movable axis generally parallel to said end edges;

c) providing cell culture media and a cell inoculate to said bag; and

d) cultivating cells in the bag with agitation provided by rocking said support around said movable axis, forming a cell suspension and supplying at least one gas via a port in said top sheet.

15. The method of claim 14, wherein said cells during cultivation express a target biomolecule, such as a protein.

16. The method of claim 14, wherein at least one baffle is tubular and joined to said bottom sheet via a cell suspension diffuser and to said top sheet via an inlet/outlet port, said method further comprising a step b′

) of connecting the inlet/outlet port on said top sheet to at least one cell separating device, such as a filter, and a pump, and in step d) circulating said cell suspension over said cell separating device or filter via said cell suspension diffuser and tubular baffle.

17. The method of claim 16, wherein said filter is a microfilter and said target biomolecule in step d) is recovered as a permeate, while a retentate is recirculated to said bag.

18. The method of claim 16, wherein said filter is an ultrafilter and in step d) said target biomolecule remains in a retentate, which is recirculated to said bag.

19. The method of claim 16, wherein in step d) said cell suspension is pumped over a retentate side of said filter by an outward stroke of a reciprocating pump and then pumped back by an inward stroke of said reciprocating pump over said retentate side into said bag via said cell suspension diffuser.

20. The method of claim 14, wherein the cultivation is performed in batch or fed-batch mode and at the end of the cultivation the cell suspension is withdrawn from the bag via a cell suspension diffuser, a tubular baffle and an outlet port and further via a cell separating device.

21. The method of claim 16, wherein in step e) said cell suspension is pumped over a retentate side of said filter in a loop back to said bag via a port on said top sheet, such as via a second tubular baffle and a second cell suspension diffuser.

22. The method of claim 14, wherein the density of viable cells is at least 40 MVC/mL, such as between 40 and 150 MVC/mL.

23. The method of claim 14 performed with an inflatable bioreactor bag.