# US 2025/0145929 A1 · Cell Expansion

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- Number: US 2025/0145929 A1
- Document ID: US20250145929A1
- Status: Published application
- Kind: A1
- Application number: 19/018,730
- Filed: 2025-01-13
- Published: 2025-05-08
- Earliest claimed: 2017-03-31
- Assignees: Terumo BCT, Inc.
- Applicants: Terumo BCT, Inc.
- Inventors: Boah VANG; Brian J. NANKERVIS; Mark E. JONES
- Web page: https://patentgrep.com/patent/US20250145929A1

## CPC

- C12M 29/10: {Perfusion}
- C12M 25/10: {Hollow fibers or tubes (hollow fiber modules in general B01D63/02)}
- C12M 35/08: {Chemical, biochemical or biological means, e.g. plasma jet, co-culture}
- C12M 41/48: {Automatic or computerized control (automatic analysis G01N35/00)}
- C12N 5/0636: {T lymphocytes}
- C12N 5/0637: {Immunosuppressive T lymphocytes, e.g. regulatory T cells or Treg}
- C12N 2501/2302: Interleukin-2 (IL-2)
- C12M: APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; {APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS}
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)

## Abstract

Embodiments described herein generally provide for expanding cells in a cell expansion system. The cells may be grown in a bioreactor, and the cells may be activated by an activator (e.g., a soluble activator complex). Nutrient and gas exchange capabilities of a closed, automated cell expansion system may allow cells to be seeded at reduced cell seeding densities, for example. Parameters of the cell growth environment may be manipulated to load the cells into a particular position in the bioreactor for the efficient exchange of nutrients and gases. System parameters may be adjusted to shear any cell colonies that may form during the expansion phase. Metabolic concentrations may be controlled to improve cell growth and viability. Cell residence in the bioreactor may be controlled. In embodiments, the cells may include T cells. In further embodiments, the cells may include T cell subpopulations, including regulatory T cells (Tregs), helper, na“i”ve, memory, or effector, for example.

## Claims

### Claim 1

A cell expansion system comprising:
a bioreactor, wherein the bioreactor comprises a first port and a second port, and wherein the first port and the second port are positioned on opposing ends of the bioreactor;
a first fluid flow path having at least opposing ends, wherein a first opposing end of the first fluid flow path is fluidly associated with the first port of the bioreactor, and a second opposing end of the first fluid flow path is fluidly associated with the second port of the bioreactor;
a fluid inlet path fluidly associated with the first fluid flow path;
a first fluid circulation path fluidly associated with the first fluid flow path and fluidly associated with the bioreactor;
a first pump to circulate fluid in the first fluid flow path;
a second pump to circulate the fluid in the first fluid circulation path;
a processor;
a memory, in communication with and readable by the processor, and
containing a series of instructions that, when executed by the processor, cause the processor to:
direct a loading of cells into the bioreactor;
direct a first feeding of the fluid to the cells in the bioreactor, wherein the fluid comprises media, and wherein the first feeding comprises:
direct the first pump to move a first portion of the fluid at a first fluid flow rate into the first port of the bioreactor, wherein the first pump moves the fluid in a first direction; and
direct the second pump to move a second portion of the fluid at a second fluid flow rate into the second port of the bioreactor, wherein the second pump moves the fluid in a second direction, wherein the second direction is opposite the first direction, and wherein the second fluid flow rate is different from the first fluid flow rate during a first time period.

### Claim 2 (depends on 1)

The cell expansion system of claim 1, wherein the first time period comprises a first step of feeding during an expansion of the cells in the bioreactor.

### Claim 3 (depends on 1)

The cell expansion system of claim 1, wherein the first fluid flow rate into the first port of the bioreactor is about 0.4 mL/min for the first time period, and wherein the second fluid flow rate into the second port of the bioreactor is about-0.2 mL/min for the first time period.

### Claim 4 (depends on 1)

The cell expansion system of claim 1, wherein the cells are exposed to an activator.

### Claim 5 (depends on 4)

The cell expansion system of claim 4, wherein the activator comprises a soluble co-sitmulatory anti-CD3/CD28/CD2 monocional antibody (mAb) T cell complex.

### Claim 6 (depends on 1)

The cell expansion system of claim 1, where in the cells comprise T cells.