BAG ASSEMBLY FOR CULTIVATION OF CELLS

• US 20170044477A1
• Filed: 04/27/2015
• Published: 02/16/2017
• Est. Priority Date: 05/28/2014
• Status: Active Grant

• Current Assignee: Cytiva Sweden AB (Danaher Corporation)
• Original Assignee: GE Healthcare Bio-Sciences AB (GE Healthcare Technologies, Inc.)
• Inventors: Gebauer, Klaus, Castan, Andreas, Estmer-Nilsson, Camilla, Falkman, Thomas, Wilen, Anders, Miller, Michael, Stankowski, Ralph
• Granted Patent: US 10,450,540 B2
• Time in Patent Office: Days
• CPC Class Codes: C12M 23/14 Bags
• CPC Class Codes: C12M 23/26 flexible flexible containers for laboratory use B01L3/505
• CPC Class Codes: C12M 23/34 Internal compartments or partitions
• CPC Class Codes: C12M 23/58 Reaction vessels connected in series or in parallel combinations of bioreactors with other apparatus, C12M43/00
• CPC Class Codes: C12M 27/16 Vibrating; Shaking; Tilting
• CPC Class Codes: C12M 41/24 inside the vessel
• CPC Class Codes: C12Q 3/00 Condition responsive control processes apparatus therefor C12M1/36; controlling or regulating in general G05

Abstract

The invention discloses a flexible bag assembly for cultivation of cells, comprising one or more bags forming a plurality of cultivation compartments, wherein a drain port in at least a first cultivation compartment is adapted to be fluidically connected with a second cultivation compartment upon opening of a valve means. It also discloses a bioreactor with the bag assembly mounted on a rocking tray and a method of cultivating cells in the assembly.

Claims

1. A flexible bag assembly for cultivation of cells, comprising one or more bags forming a plurality of cultivation compartments, wherein a drain port in at least a first cultivation compartment is adapted to be fluidically connected with a second cultivation compartment upon opening of a valve means.

2. The flexible bag assembly of claim 1, wherein said second cultivation compartment has a volume of at least 120%, such as at least 150% of the volume of the first cultivation compartment.

3. The flexible bag assembly of claim 1, comprising at least three cultivation compartments and wherein a drain port in the second cultivation compartment is adapted to be fluidically connected with a third cultivation compartment upon opening of a valve means.

4. The flexible bag assembly of claim 3, wherein said third cultivation compartment has a volume of at least 120%, such as at least 150% of the volume of the second cultivation compartment.

5. The flexible bag assembly of claim 1, wherein at least one cultivation compartment, such as the first cultivation compartment, is a rigid or semirigid vessel.

6. The flexible bag assembly of claim 1, wherein said valve means is a valve or a clamp.

7. The flexible bag assembly of claim 6, wherein said valve means is a pinch valve or a pinch clamp.

8. The flexible bag assembly of claim 7, wherein said pinch valve or pinch clamp is adapted for application on tubing or conduits of up to 15 mm outer diameter.

9. The flexible bag assembly of any preceding claim 1, wherein at least one of said drain ports is a dip tube.

10. The flexible bag assembly of claim 1, comprising at least two bags, such as at least three bags, connected by tubing.

11. The flexible bag assembly of claim 10, wherein said at least two bags are adapted to be placed beside each other on a tray.

12. The flexible bag assembly of claim 10, wherein said at least two bags are adapted to be stacked on a tray or a stack of trays.

13. The flexible bag assembly of claim 1, comprising at least one multicompartment bag with cultivation compartments connected or connectable via internal conduits.

14. The flexible bag assembly of claim 1, comprising at least one multicompartment bag with cultivation compartments connectable via at least one breakable seal.

15. The flexible bag assembly of claim 1, comprising at least one bag comprising at least one temperature control compartment.

16. The flexible bag assembly of claim 15, wherein said at least one bag comprises a top film, a bottom film and a delimiting film, wherein the delimiting film delimits the at least one temperature control compartment from at least one cultivation compartments, such as from at least said first cultivation compartment.

17. The flexible bag assembly of claim 16, wherein said temperature control compartment comprises a thermostating fluid inlet and optionally a thermostating fluid outlet.

18. The flexible bag assembly of claim 17, wherein the volume of said first cultivation compartment is less than 1 L, such as 50-250 mL, and/or the volume of said temperature control compartment is at least 10 times the volume of said first cultivation compartment.

19. The flexible bag assembly of claim 1, adapted for cell cultivation on a rocking tray platform.

20. The flexible bag assembly of claim 1, wherein each cultivation compartment comprises a gas inlet and a gas outlet.

21. The flexible bag assembly of claim 1, supplied presterilized, such as by radiation sterilization.

22. A bioreactor assembly, comprising the flexible bag assembly of claim 1 mounted on a tray, or a stack of trays, adapted to rock back and forth around at least one axis.

23. The bioreactor assembly of claim 22, comprising a cell culture in at least one of said cultivation compartments.

24. A method of cultivating cells, comprising the steps of:

a) providing the bioreactor assembly of claim 22;

b) introducing culture media and cells in said first cultivation compartment;

c) cultivating the cells in said first cultivation compartment to provide a first cell culture;

d) opening a first valve means to fluidically connect said first cultivation compartment with said second cultivation compartment;

e) transferring said first cell culture to said second cultivation compartment;

f) introducing media to said second cultivation compartment, and;

g) cultivating the cells in said second cultivation compartment to provide a second cell culture.

25. The method of claim 24, further comprising a step e′

), after step e), of closing the first valve means.

26. The method of claim 25, further comprising the steps of:

h) opening a second valve means to fluidically connect said second cultivation compartment with a third cultivation compartment;

i) transferring said second cell culture to said third cultivation compartment;

j) introducing media to said third cultivation compartment, and;

k) cultivating the cells in said third cultivation compartment to provide a third cell culture.

27. The method of claim 26, further comprising a step i′

), after step i), of closing said (second) valve means.

28. A flexible bag assembly for cultivation of cells, comprising at least one bag with at least one cultivation compartment and at least one temperature control compartment.

29. The flexible bag assembly of claim 28, wherein said at least one bag comprises a top film, a bottom film and a delimiting film, wherein the delimiting film delimits the at least one temperature control compartment from at least one cultivation compartment, such as each cultivation compartment.

30. The flexible bag assembly of claim 29, wherein at least one cultivation compartment, such as each cultivation compartment, has a volume of less than 1 L, such as 50-250 mL, and/or the volume of said temperature control compartment is at least 10 times the volume of at least one cultivation compartment, such as each cultivation compartment.

31. The flexible bag assembly of claim 30, wherein said temperature control compartment comprises an inlet and an outlet for a thermostating fluid.

32. A bioreactor assembly comprising:

a) at least one bag or bottle with at least one cultivation compartment;

b) at least one flexible temperature control compartment comprising a thermostating fluid, and;

c) a tray adapted to rock back and forth around at least one axis,

wherein said at least one flexible temperature control compartment is mounted on said tray and said at least one bag or bottle is located on said at least one flexible temperature control compartment.

33. The bioreactor assembly of claim 32, wherein said at least one flexible temperature control compartment forms part of said at least one bag.

34. The bioreactor assembly of claim 32, wherein said at least one flexible temperature control compartment forms part of a second bag separate from said at least one bag.

35. A method of cultivating cells, comprising the steps of:

a) providing the bioreactor assembly of claim 34;

b) introducing culture media and cells in said at least one cultivation compartment;

c) cultivating the cells in said at least one cultivation compartment.

36. The method of claim 35, wherein the temperature in step c) is maintained within a range of a target temperature /−

2°

C., such as /−

1°

C. or /−

0.5°

C.

37. The method of claim 36, wherein the thermostating fluid is circulated through said flexible temperature control compartment.

38. The method of claim 37, wherein said at least one cultivation compartment comprises a temperature sensor which is electrically or electromagnetically connected to a control unit and the temperature of said thermostating fluid is controlled by the control unit using a feedback loop.