Coating a bioreactor in a cell expansion system

Patent metadata

Field	Value
Publication number	US-12077739-B2
Application number	US-202318169961-A
Country	US
Kind code	B2
Filing date	Feb 16, 2023
Priority date	Jun 7, 2016
Publication date	Sep 3, 2024
Grant date	Sep 3, 2024

Abstract

Embodiments described herein generally provide for the expansion of cells in a cell expansion system using an active promotion of a coating agent(s) to a cell growth surface in some embodiments. A coating agent may be applied to a surface, such as the cell growth surface of a hollow fiber in a bioreactor, by controlling the movement of a fluid in which a coating agent is suspended, by changing flow rates, by changing flow directions, by rotation of the bioreactor, and/or combinations thereof.

First claim

What is claimed is:

1. A method of growing cells in a bioreactor, the method comprising:
circulating a first fluid at a first flow rate through a bioreactor of a cell expansion system, wherein the first fluid comprises a reagent;
circulating the first fluid for a first period of time to allow at least a first portion of the reagent to coat the bioreactor;
after the first period of time, circulating the first fluid at a second flow rate slower than the first flow rate through the bioreactor of the cell expansion system for a second period of time to allow a second portion of the reagent to coat the bioreactor;
circulating a second fluid through the bioreactor to remove a third portion of the reagent from the bioreactor;
circulating a third fluid at a third flow rate through a bioreactor of a cell expansion system, wherein the third fluid comprises a plurality of cells;
reducing circulation of the third fluid from the third flow rate;
maintaining the bioreactor in a first orientation for a third predetermined period of time to allow at least a portion of the plurality of cells to settle;
after the third predetermined period of time, rotating the bioreactor to a second orientation that is about 180 degrees from the first orientation;
after the rotating, maintaining the bioreactor in the second orientation for a fourth period of time to allow at least a second portion of the plurality of cells to settle; and
expanding at least the first and the second portion of cells in the bioreactor.

2. The method of claim 1 , further comprising:
after the third predetermined period of time and before the maintaining the bioreactor in the second orientation for a fourth period of time:
circulating the third fluid at a fourth flow rate through the bioreactor, wherein the fourth flow rate is slower than the third flow rate; and
reducing the circulation from the fourth flow rate; and

maintaining the bioreactor in the second orientation for the fourth period of time.

3. The method of claim 2 , further comprising:
after the fourth predetermined period of time:
circulating the third fluid at a fifth flow rate through the bioreactor, wherein the fifth flow rate is slower than the fourth flow rate;
reducing a rate of circulation of the third fluid from the fifth flow rate; and
maintaining the bioreactor in the second orientation for a fifth predetermined period of time to allow at least a third portion of the plurality of cells to settle.

4. The method of claim 3 , further comprising:
after the fifth predetermined period of time:
circulating fluid at a sixth flow rate through the bioreactor, wherein the sixth flow rate is slower than the fifth flow rate; and
reducing circulation of fluid from the sixth flow rate; and
maintaining the bioreactor in the second orientation for a sixth predetermined period of time to allow at least a fourth portion of the plurality of cells to settle.

5. The method of claim 4 , wherein the third period of time, the fourth period of time, the fifth period of time, and the sixth period of time are the same.

6. The method of claim 5 , wherein the third period of time and the fourth period of time are equal in duration.

7. The method of claim 6 , wherein the first period of time is less than about 10 minutes.

8. The method of claim 7 , wherein the bioreactor comprises a hollow fiber membrane.

9. The method of claim 1 , further comprising:
after the second predetermined period of time and before expanding, rotating the bioreactor 180 degrees to the first orientation.

Assignees

• Terumo Bct Inc

Inventors

• Frank Nathan D
• Nankervis Brian J
• Hlavinka Dennis J
• Dilorenzo Thomas G

Classifications

• C12M1/36

  including condition or time responsive control, e.g. automatically controlled fermentors (controlling or regulating in general G05) · CPC title

• C12M29/10

  Perfusion · CPC title

• C12N5/0663

  Bone marrow mesenchymal stem cells (BM-MSC) · CPC title

• C12M25/12

  the culture medium flowing outside the fiber or tube · CPC title

• C12M29/18

  External loop; Means for reintroduction of fermented biomass or liquid percolate (loop type reactors for chemical or physical processes B01J19/2435) · CPC title