Vertical wheel bioreactors

• US 9,453,194 B2
• Filed: 07/28/2014
• Issued: 09/27/2016
• Est. Priority Date: 10/26/2005
• Status: Active Grant

• Current Assignee: PBS Biotech, Inc.
• Original Assignee: PBS Biotech, Inc.
• Inventors: Zeikus, J. Gregory
• Primary Examiner(s): SOOHOO, TONY GLEN
• Application Number:
  US14/444,695
• Publication Number:
  US 20140335597A1
• Time in Patent Office: 792 Days
• US Class Current: 1/1
• CPC Class Codes: B01F 23/233 using driven stirrers with completely immersed stirring elements
• CPC Class Codes: B01F 23/23351 the gas moving along the axis of rotation
• CPC Class Codes: B01F 23/23362 the gas being introduced under the stirrer
• CPC Class Codes: B01F 27/073 with stirring elements moving with respect to the stirrer shaft, e.g. floating or comprising contracting chambers
• CPC Class Codes: B01F 27/11252 paddle wheels
• CPC Class Codes: B01F 27/55 with stirrers driven by the moving material
• CPC Class Codes: B01F 27/70 with paddles, blades or arms
• CPC Class Codes: B01F 27/91 with propellers
• CPC Class Codes: C12M 23/28 disposable or single use
• CPC Class Codes: C12M 23/56 Floating elements
• CPC Class Codes: C12M 27/00 Means for mixing, agitating or circulating fluids in the vessel by introduction of gas C12M29/06, C12M29/14, mixing in general or mixers per se B01F; mixing in apparatus for chemical or physical processes B01J
• CPC Class Codes: C12M 27/06 with horizontal or inclined stirrer shaft or axis
• CPC Class Codes: C12M 37/00 Means for sterilizing, maintaining sterile conditions or avoiding chemical or biological contamination C12M23/38 takes precedence; filtration in general and filters per se B01D24/00-B01D41/00; autoclaves B01J3/04; treatment of microorganisms with electrical or wave energy C12N13/00
• CPC Class Codes: C12M 41/12 of temperature controlling the temperature of chemical or physical processes B01J19/0013, heating or cooling apparatus for laboratory use B01L7/00
• CPC Class Codes: C12M 41/42 of agitation speed

Abstract

A pneumatic bioreactor includes a vessel containing a fluid to be mixed and at least one mixing device driven by gas pressure. A first embodiment includes a floating impeller that rises and falls in the fluid as gas bubbles carry it upward to the surface where the gas is then vented, permitting the impeller to sink in the fluid. The floating impeller may be tethered to a second impeller with a flexible member and pulley. The mixing speed is controlled with electromagnets in the vessel acting upon magnetic material in the impeller or its guides. In another embodiment, floating pistons mix the fluid, pushing it through a mixing plate with one or more apertures. In a third embodiment, the mixing device is a rotating drum with bubble-catching blades and rotating mixing plates with apertures. The top of the vessel for these mixers may include a closed top and sterile filters.

Claims

1. A method for performing a cell culture growth process, comprising:

providing a containment vessel for holding a cell culture medium, the containment vessel comprising a generally semi-cylindrical bottom wall defining a semi-cylindrical concavity therewithin, the containment vessel having a rotatable mixing wheel mounted within for rotation about a horizontal axis approximately coincident with an axis of curvature of the semi-cylindrical bottom wall, the mixing wheel having a rotational diameter that extends into close proximity with the semi-cylindrical bottom wall;

sterilizing the containment vessel and the mixing wheel so that at least the interior of the containment vessel and the mixing wheel are sterile;

providing a cell culture medium within the containment vessel;

providing nutrients within the containment vessel to support growth of a cell culture within the cell culture medium;

enclosing the containment vessel such that the cell culture medium is not exposed to the surrounding environment; and

rotating the mixing wheel about the horizontal axis to mix the contents of the containment vessel.

2. The method of claim 1, further comprising providing a temperature control sheet arranged in contact with the containment vessel bottom wall for controlling the temperature of the interior of the containment vessel and support growth of the cell culture.

3. The method of claim 1, further including a rigid housing into which the containment vessel fits, the housing defining a semi-cylindrical lower section and a lower portion of the containment vessel being formed of a material that is non-structural such that the semi-cylindrical lower section of the housing structurally supports the semi-cylindrical bottom wall of the containment vessel.

4. The method of claim 3, wherein a vessel top extends outwardly of containment vessel sides and rests on upper edges of the housing so as to suspend the containment vessel within the housing.

5. The method of claim 3, wherein the containment vessel is primarily formed of a thin wall film that is non-structural.

6. The method of claim 1, further including an impeller defined within an outer rim on the mixing wheel configured to generate axial flow of at least a portion of the contents of the containment vessel upon rotation of the mixing wheel.

7. The method of claim 1, further including a paddle that extends axially outward from the mixing wheel to mix liquid between the mixing wheel and inner sides of the containment vessel.

8. The method of claim 1, further including introducing air bubbles into the lower portion of the containment vessel to pneumatically rotate the mixing wheel.

9. A method for performing a cell culture growth process, comprising:

providing a containment vessel for holding a cell culture medium, the containment vessel comprising a mixing chamber therewithin and having therein a rotatable mixing wheel mounted for rotation about a horizontal axis, the mixing wheel substantially filling a lower portion of the mixing chamber, the containment vessel having a vessel top and a lower portion formed of a material that is non-structural;

sterilizing the containment vessel and the mixing wheel so that at least the interior of the containment vessel and the mixing wheel are sterile;

suspending the containment vessel within a rigid housing having upper edges and a bottom wall on which is supported the non-structural lower portion of the containment vessel, and further wherein the housing has side walls that closely surround and support peripheral walls of the containment vessel;

providing a cell culture medium within the containment vessel;

providing nutrients within the containment vessel to support growth of a cell culture within the cell culture medium;

enclosing the containment vessel such that the cell culture medium is not exposed to the surrounding environment;

rotating the mixing wheel about the horizontal axis to mix the contents of the containment vessel;

removing the containment vessel from the housing after a single use of growing a cell culture;

removing the containment vessel and mixing wheel from within the housing; and

suspending a second containment vessel and mixing wheel within the housing and performing a cell culture growth process.

10. The method of claim 9, wherein the containment vessel comprises a generally semi-cylindrical bottom wall defining a semi-cylindrical lower portion of the mixing chamber, the horizontal axis being approximately coincident with an axis of curvature of the semi-cylindrical bottom wall and the mixing wheel having a rotational diameter that extends into close proximity with the semi-cylindrical bottom wall.

11. The method of claim 10, wherein the housing bottom wall is semi-cylindrical in shape and supports the semi-cylindrical bottom wall of the containment vessel.

12. The method of claim 9, further including an impeller defined within an outer rim on the mixing wheel configured to generate axial flow of at least a portion of the contents of the containment vessel upon rotation of the mixing wheel.

13. The method of claim 9, further including a paddle that extends axially outward from the mixing wheel to mix liquid between the mixing wheel and inner sides of the containment vessel.

14. The method of claim 9, further including introducing air bubbles into the lower portion of the containment vessel to pneumatically rotate the mixing wheel.

15. A method for performing a cell culture growth process, comprising:

providing a containment vessel for holding a cell culture medium, the containment vessel comprising a mixing chamber therewithin and having therein a rotatable mixing wheel mounted for rotation about a horizontal axis, the mixing wheel substantially filling a lower portion of the mixing chamber adjacent a bottom wall, the mixing wheel having an outer rim connected to an axle with spokes and having an impeller located radially within the outer rim having paddles that extend outward from the outer rim for urging flow off of the bottom wall upon rotation of the mixing wheel;

sterilizing the containment vessel and the mixing wheel so that at least the interior of the containment vessel and the mixing wheel are sterile;

providing a cell culture medium within the containment vessel;

providing nutrients within the containment vessel to support growth of a cell culture within the cell culture medium;

enclosing the containment vessel such that the cell culture medium is not exposed to the surrounding environment; and

rotating the mixing wheel about the horizontal axis to mix the contents of the containment vessel.

16. The method of claim 15, wherein the bottom wall of the containment vessel is generally semi-cylindrical thus defining semi-cylindrical lower portion of the mixing chamber, the horizontal axis being approximately coincident with an axis of curvature of the semi-cylindrical bottom wall and the mixing wheel having a rotational diameter that extends the paddles into close proximity with the semi-cylindrical bottom wall.

17. The method of claim 15, further comprising providing a temperature control sheet arranged in contact with the containment vessel bottom wall for controlling the temperature of the interior of the containment vessel and support growth of the cell culture.

18. The method of claim 15, wherein the mixing wheel includes two parallel plates axially spaced from one another and a plurality of mixing elements evenly arranged therebetween around an outer periphery of the mixing wheel.

19. The method of claim 17, wherein the mixing wheel further includes a paddle that extends axially outward from the parallel plates to mix liquid between the mixing wheel and inner sides of the containment vessel.

20. The method of claim 15, further including introducing air bubbles into the lower portion of the containment vessel to pneumatically rotate the mixing wheel.