Expanding cells in a bioreactor

• US 10,633,625 B2
• Filed: 11/14/2014
• Issued: 04/28/2020
• Est. Priority Date: 11/16/2013
• Status: Active Grant

• Current Assignee: Terumo BCT, Inc. (Terumo Corporation)
• Original Assignee: Terumo BCT, Inc. (Terumo Corporation)
• Inventors: Nankervis, Brian J., Jones, Mark E.
• Primary Examiner(s): Singh, Satyendra K
• Application Number:
  US14/542,304
• Publication Number:
  US 20150140654A1
• Time in Patent Office: 1,992 Days
• CPC Class Codes: C12M 23/50 Means for positioning or orientating the apparatus C12M41/08 takes precedence
• CPC Class Codes: C12M 25/10 Hollow fibers or tubes hollow fiber modules in general B01D63/02
• CPC Class Codes: C12M 25/12 the culture medium flowing outside the fiber or tube
• CPC Class Codes: C12M 27/10 Rotating vessel
• CPC Class Codes: C12M 29/10 Perfusion
• CPC Class Codes: C12M 29/16 Hollow fibers hollow fiber modules in general B01D63/02
• CPC Class Codes: C12M 29/18 External loop; Means for reintroduction of fermented biomass or liquid percolate loop type reactors for chemical or physical processes B01J19/2435
• CPC Class Codes: C12M 41/48 Automatic or computerized control automatic analysis G01N35/00
• CPC Class Codes: C12N 5/0663 Bone marrow mesenchymal stem cells (BM-MSC)

Abstract

Described are embodiments for expanding cells in a bioreactor. In one embodiment, methods are provided that distribute cells throughout the bioreactor and attach cells to specific portions of a bioreactor to improve the expansion of the cells in the bioreactor. Embodiments may be implemented on a cell expansion system configured to load, distribute, attach and expand cells.

Claims

1. A method of growing cells in a cell expansion system, the method comprising:

circulating fluid, with a pump, through a bioreactor comprising a hollow fiber membrane with a plurality of hollow fibers, wherein the fluid comprises a plurality of cells and the fluid enters the bioreactor through a first inlet and exits the bioreactor through a first outlet;

reducing the rate of circulating fluid by reducing a flow rate of the pump;

maintaining the bioreactor in a first horizontal orientation for a first predetermined period of time to allow at least a first portion of the plurality of cells to settle and attach to a first portion of at least one of the plurality of hollow fibers;

after the first predetermined period of time, rotating, with a rotational device, the bioreactor to a second horizontal orientation that is about 180 degrees from the first horizontal orientation; and

after the rotating, expanding at least the first portion of the plurality of cells attached to the first portion of the at least one of the plurality of hollow fibers while the bioreactor is oriented in the second horizontal orientation by feeding the portion of cells and growing the portion of cells downward in the bioreactor under an influence of gravity toward a portion of the at least one of the plurality of hollow fibers that does not have attached cells.

2. The method of claim 1, wherein the first predetermined period of time is long enough to allow the portion of cells to attach and expand.

3. The method of claim 2, wherein the first predetermined period of time is greater than about 10 hours.

4. The method of claim 1, wherein the first predetermined period of time is less than about 8 minutes.

5. The method of claim 4, wherein the first predetermined period of time is about 5 minutes.

6. The method of claim 1, further comprising:

after the first predetermined period of time and before rotating the bioreactor to a second horizontal orientation;

rotating the bioreactor to a first vertical orientation and maintaining the bioreactor in the first vertical orientation for a second predetermined period of time; and

after the second predetermined period of time, rotating the bioreactor to a second vertical orientation that is about 180 degrees from the first vertical orientation and maintaining the bioreactor in the second vertical orientation for a third predetermined period of time.

7. The method of claim 6, wherein the second predetermined period of time is of sufficient duration to allow the portion of cells to expand.

8. The method of claim 7, wherein the third predetermined period of time is equal in duration to the second period of time.

9. The method of claim 7, wherein the second predetermined period of time is at least about 10 hours.

10. The method of claim 1, wherein the plurality of cells comprises stem cells.

11. The method of claim 10, wherein the stem cells comprise mesenchymal stem cells.

12. The method of claim 1, wherein the plurality of cells comprises one or more cells from the group consisting of:

fibroblasts, keratinocytes, progenitor, endothelial, or combinations thereof.

13. The method of claim 1, wherein the first portion comprises an inside surface of the at least one of the plurality of hollow fibers.

14. The method of claim 13, wherein the second portion comprises a different portion of the inside surface of the at least one of the plurality of hollow fibers.

15. The method of claim 1, wherein the first predetermined period of time is long enough to allow the portion of cells to attach.