JP-2026514488-A - A method for removing solutes without losing suspended cells during the cell proliferation process.

Abstract

[Solution] A method for removing solute while retaining suspended cells in a bioreactor comprises the steps of positioning a suspension containing cells within a predetermined area of the bioreactor and generating a flow rate in the suspension within the bioreactor, wherein the flow rate is selected such that the cells are retained in the bioreactor while the solute exits the bioreactor. [Selection Diagram] None

Inventors

• ラヴィンカ、デニス ジェイ．

Assignees

• テルモ ビーシーティー、インコーポレーテッド

Dates

Publication Date

20260511

Application Date

20240418

Priority Date

20240417

Claims (20)

1. A method for removing solutes while retaining suspended cells in a bioreactor, the method being: The steps include positioning a suspension containing cells and solute within a predetermined area of the bioreactor, A step of generating a flow rate in the suspension within the bioreactor, wherein the flow rate is selected such that the cells are maintained within the bioreactor while the solute exits the bioreactor. Having, method.
2. In the method described in claim 1, The step of positioning the suspension is: The steps include generating a first flow at the first end of the bioreactor, The steps include generating a second flow at the second end of the bioreactor, It has, The first flow and the second flow work together to adjust the first axial flow from the first end and the second axial flow from the second end so that the suspension is positioned within the predetermined area of the bioreactor. method.
3. In the method described in claim 2, The first flow has a first flow rate, and the second flow has a second flow rate that is different from the first flow rate. method.
4. In the method described in claim 3, The second flow rate is smaller than the first flow rate. method.
5. In the method described in claim 4, The second flow rate is less than half of the first flow rate. method.
6. In the method described in claim 2, The step of positioning the suspension further includes: The step of positioning the bioreactor in a horizontal or substantially horizontal position such that the suspension settles on or near the wall defining the bioreactor, method.
7. In the method described in claim 6, After the step of positioning the suspension and before the step of generating the flow rate in the suspension, the method further: The process includes a step of stopping all flow through the bioreactor. method.
8. In the method described in claim 7, After the step of stopping all flow through the bioreactor and before the step of generating the flow rate in the suspension, the method further: The step of setting a flow valve to allow the flow to exit the bioreactor, method.
9. In the method described in claim 7, Prior to the step of generating the flow rate in the suspension, the method further: The step of positioning the bioreactor at an angle of 45 degrees or approximately 45 degrees with respect to the horizontal, method.
10. In the method described in claim 1, The predetermined region of the bioreactor is a region located in the center between the first end of the bioreactor and the second end opposite to the first end. method.
11. In the method according to claim 10, The predetermined region is perpendicular to the long axis of the bioreactor. method.
12. A method for removing solutes while retaining suspended cells in a bioreactor, the method being: The steps include positioning a suspension containing cells and solute within a predetermined area of the bioreactor, The steps include: positioning the bioreactor so as to define its upper and lower ends; The steps include preventing flow from occurring from the upper end of the bioreactor, The steps include: introducing the culture medium into the lower end of the bioreactor; The steps include enabling flow from the upper end of the bioreactor, The steps include preventing flow from occurring from the lower end of the bioreactor, The steps include: flowing a fluid across the bioreactor to move the solute out of the bioreactor and retain the cells within the bioreactor; Having, method.
13. In the method according to claim 12, The predetermined region of the bioreactor is a region located in the center between the first end of the bioreactor and the second end opposite to the first end. method.
14. In the method according to claim 12, The predetermined region is perpendicular to the long axis of the bioreactor. method.
15. A method for removing solutes while retaining suspended cells in a bioreactor, the method being: The steps include: positioning the bioreactor at a certain angle to the horizontal so as to define the upper and lower ends of the bioreactor; The steps include positioning a suspension containing cells and solute in a predetermined location within the bioreactor, Steps include starting the flow at a circulating flow rate, The steps include: starting the flow at a pre-selected net ultrafiltration flow rate, Having, method.
16. In the method described in claim 15, The step of positioning the suspension is: The steps include generating a first flow at the lower end of the bioreactor, The steps include generating a second flow at the upper end of the bioreactor, It has, The first flow and the second flow work together to adjust the first axial flow from the first end and the second axial flow from the second end so that the suspension is positioned in the predetermined location within the bioreactor. method.
17. In the method according to claim 16, The first flow has a first flow rate, and the second flow has a second flow rate that is different from the first flow rate. method.
18. In the method described in claim 17, The second flow rate is smaller than the first flow rate. method.
19. In the method described in claim 18, The second flow rate is less than half of the first flow rate. method.
20. In the method described in claim 15, After the step of positioning the suspension in the predetermined location within the bioreactor and before the step of starting the flow at the circulating flow rate, the method further: It has a step to stop all flow, method.

Description

(Cross-reference of related applications) This application claims priority to U.S. Patent Application No. 18/637,782, filed on 17 April 2024, and also claims the benefit of U.S. Provisional Patent Application No. 63/461,989, filed on 26 April 2023. The entire disclosure of the above application is incorporated herein by reference. This disclosure relates to a method for removing high molecular weight solutes without losing suspended cells during a cell proliferation process. This section provides background information related to this disclosure that is not necessarily prior art. A hollow fiber cell proliferation system, also called a cell proliferation system and/or hollow fiber bioreactor, is a cell culture system comprising one or more detachable bioreactor cartridges used to proliferate and differentiate cells, including both adherent and non-adherent cell types. The bioreactor cartridge comprises a hollow fiber membrane containing a plurality of semipermeable hollow fibers. The space inside the hollow fibers (i.e., the lumen) defines the inner capillary space, while the space outside the hollow fibers defines the outer capillary space. Depending on the permeability of the hollow fiber walls, high molecular weight solutes (e.g., with a molecular weight of approximately 20,000 Daltons or more) from the culture medium and/or the growing cell population may not be able to easily diffuse through the hollow fiber walls, resulting in the undesirable gradual accumulation of solutes (e.g., proteins, cytokines, and/or waste products) in the inner capillary space (or, alternatively, in addition, the outer capillary space). Solute accumulation has undesirable effects on cell viability, proliferation, and/or phenotype. Figure 1 shows an example of a cell proliferation system having a bioreactor according to at least one exemplary embodiment.Figure 2 is a diagram of an example bioreactor showing a circulation path through the bioreactor, which is incorporated into a cell proliferation system similar to the cell proliferation system illustrated in Figure 1, according to at least one exemplary embodiment.Figure 3 shows an example of a rocking device configured to be used in conjunction with a cell proliferation system similar to the cell proliferation system shown in Figure 1 to operate a bioreactor similar to the bioreactor in Figure 2, according to at least one exemplary embodiment.Figure 4 is a schematic diagram showing an example of a flow path in a cell proliferation system similar to the cell proliferation system shown in Figure 1, according to at least one exemplary embodiment.Figure 5 is a flowchart illustrating an exemplary method for removing solute from the capillary inner space (or, alternatively, or in addition to, the capillary outer space) of a bioreactor similar to the bioreactor shown in Figure 2, according to at least one exemplary embodiment.Figure 6 is a simplified diagram illustrating the exemplary method shown in Figure 5, applied to a single hollow fiber of a bioreactor according to at least one exemplary embodiment.Figure 7 is a flowchart illustrating another exemplary method for removing solute from the capillary inner space (or, alternatively, or in addition to, the capillary outer space) of a bioreactor similar to the bioreactor shown in Figure 2, according to at least one exemplary embodiment.Figure 8 is a simplified diagram illustrating the exemplary method shown in Figure 7, applied to a single hollow fiber of a bioreactor according to at least one exemplary embodiment.Figure 9 is a flowchart illustrating another exemplary method for removing solute from the capillary inner space (or, alternatively, or in addition to, the capillary outer space) of a bioreactor similar to the bioreactor shown in Figure 2, according to at least one exemplary embodiment.Figure 10 is a simplified diagram illustrating the exemplary method shown in Figure 9, applied to a single hollow fiber of a bioreactor according to at least one exemplary embodiment.Figure 11 is a flowchart illustrating another exemplary method for removing solute from the capillary inner space (or, alternatively, or in addition to, the capillary outer space) of a bioreactor similar to the bioreactor shown in Figure 2, according to at least one exemplary embodiment.Figure 12A is a simplified diagram illustrating the exemplary method shown in Figure 11, applied to a single hollow fiber of a bioreactor according to at least one exemplary embodiment.Figure 12B is a simplified diagram illustrating the exemplary method shown in Figure 11, applied to a single hollow fiber of a bioreactor according to at least one exemplary embodiment. The corresponding reference numerals indicate the corresponding parts in the above diagram. Exemplary embodiments are described in more detail with reference to the accompanying drawings. Exemplary embodiments are provided to those skilled in the art to ensure that the disclosure is complete and its scope fully conve