Method of forming a collapsible bag using a mold and mandrel

1. A bioreactor comprising:

a collapsible bag configured to be received in a reusable rigid support structure, the collapsible bag having an interior configured to receive a volume of liquid, the collapsible bag including an impeller support having a rotatable impeller;

a first sparger connected to the collapsible bag, the first sparger having a port configured for attachment to tubing in fluid communication with a source of gas; and

a second sparger connected to the collapsible bag, the second sparger having a port configured for attachment to tubing in fluid communication with a source of gas;

wherein the first and second spargers are formed in the impeller support of the collapsible bag;

wherein the first sparger has a first aperture size, and wherein the second sparger has a second aperture size that is different from the first aperture size;

wherein the impeller support has a first portion configured to receive and/or support a shaft of the impeller and a second portion adjacent to the first portion, the second portion being thinner than the first portion; and

wherein the first sparger and second sparger are formed in the first portion of the impeller support.

2. The bioreactor of claim 1 wherein the first portion has a size that is substantially the same as a diameter of the impeller.

3. The bioreactor of claim 1 , further comprising:

a control system operatively associated with the first and second spargers and configured to operate the first and second spargers independently of each other.

4. The bioreactor of claim 1 , wherein the vessel comprises greater than 10 spargers, each connected to the collapsible bag.

5. The bioreactor of claim 1 , wherein the vessel comprises greater than 20 spargers, each connected to the collapsible bag.

6. The bioreactor of claim 1 , wherein collapsible bag has a volume of at least 2 liters.

7. The bioreactor of claim 1 , wherein the first sparger and/or the second sparger are in fluid communication with a source of N 2 , O 2 , CO 2 , NH 3 , and/or air.

8. The bioreactor of claim 7 , wherein the first sparger is in fluid communication with a source of a gas comprising air and the second sparger is in fluid communication with a source of a gas comprising air supplemented with O 2 and/or N 2 .

9. The bioreactor of claim 1 , wherein first aperture size is larger than the second aperture size.

10. The bioreactor of claim 1 , wherein the first aperture size is between 0.1 mm and 10 mm.

11. The bioreactor of claim 1 , wherein the first aperture size is between 100 microns and 10 mm.

12. The bioreactor of claim 1 , wherein the first aperture size is between 500 microns and 3 mm.

13. The bioreactor of claim 1 , wherein the first aperture size is between 500 microns and 1 mm.

14. The bioreactor of claim 1 , wherein the second aperture size is between 0.1 microns and 200 microns.

15. The bioreactor of claim 1 , wherein the second aperture size is between 0.1 microns and 100 microns.

16. The bioreactor of claim 1 , wherein the second aperture size is between 0.1 microns and 50 microns.

17. The bioreactor of claim 1 , wherein the second aperture size is between 100 microns and 200 microns.

18. The bioreactor of claim 1 , wherein:

the first sparger and the second sparger are connected to a bottom of the collapsible bag.

19. The bioreactor of claim 1 , wherein:

the first and second spargers each include tubing configured for fluid communication with the source of gas and a sparging element having a plurality of apertures.

20. The bioreactor of claim 1 , wherein the collapsible bag is configured to receive a sensor.