Methods of cell culture for adoptive cell therapy

• US 8,809,050 B2
• Filed: 12/08/2010
• Issued: 08/19/2014
• Est. Priority Date: 12/08/2009
• Status: Active Grant

• Current Assignee: Baylor College of Medicine (Baylor College), Wilson Wolf Manufacturing Corporation
• Original Assignee: Wilson Wolf Manufacturing Corporation
• Inventors: Vera, Juan F., Leen, Ann M., Wilson, John R., Rooney, Cliona M
• Primary Examiner(s): Noakes, Suzanne M
• Assistant Examiner(s): CHONG, STEPHEN M
• Application Number:
  US12/963,597
• Publication Number:
  US 20110136228A1
• Time in Patent Office: 1,350 Days
• Field of Search: None
• US Class Current: 435/325
• CPC Class Codes: A61K 39/4611 T-cells, e.g. tumor infiltrating lymphocytes [TIL], lymphokine-activated killer cells [LAK] or regulatory T cells [Treg]
• CPC Class Codes: A61K 39/464838 Viral antigens
• CPC Class Codes: C12N 2502/11 blood or immune system cells
• CPC Class Codes: C12N 5/0636 T lymphocytes

Abstract

An improved method of culturing cells for cell therapy applications that includes growing desired cells in the presence of antigen-presenting cells and/or feeder cells and with medium volume to surface area ratio of up to 1 ml/cm² if the growth surface is not comprised of gas permeable material and up to 2 ml/cm² if the growth surface is comprised of gas permeable material. The desired cells are at a surface density of less than 0.5×10⁶ cells/cm² at the onset of a production cycle, and the surface density of the desired cells plus the surface density of the antigen presenting cells and/or feeder cells are at least about 1.25×10⁵ cells/cm².

Claims

1. An improved method of producing cells comprising:

growing activated antigen-specific T cells in the presence of irradiated antigen-presenting cells-on a growth surface and with a medium volume to surface area ratio of up to 1 ml/cm²;

wherein said surface density of the antigen-specific T cells plus the surface density of said antigen presenting cells is between 128,906 and 1,125,000 cells/cm² and the surface density of the antigen-specific T cells is less than 250,000 cells/cm² and the combined number of said antigen presenting cells and antigen-specific T cells is less than 500,000 cells/cm² at the onset of a production cycle.

2. The method of claim 1 wherein growing antigen-specific T cells includes more than one production cycle.

3. The method of claim 1 wherein said antigen-specific T cells are Epstein Barr Virus Cytotoxic T lymphocytes.

4. The method of claim 1 wherein the method is a static cell culture method.

5. The method of claim 1 wherein the growth surface resides in a standard 24 well plate.

6. A method of producing cells comprising:

Growing activated antigen specific T cells on a growth surface wherein said antigen specific T cells reside at a surface density of between 3.1×

10⁴ and 6.2×

10⁴ antigen specific T cells per cm² of growth surface area and in the presence of antigen presenting cells residing at ratio of between 2 and 32 times the number of antigen specific T cells.

7. The method of claim 6 including more than one production cycle.

8. The method of claim 6 wherein said antigen specific T cells have antigen specificity to a virus.

9. The method of claim 8 wherein said virus is Epstein Barr Virus.

10. The method of claim 6 wherein said antigen presenting cells are lymphoblastoid cells.

11. The method of claim 6 wherein the method is a static cell culture method.

12. The method of claim 6 wherein said growth surface is gas permeable.

13. The method of claim 12 wherein the highest level of said media volume is beyond 2.0 cm from the lowest level of said media volume.

14. The method of claim 6 wherein the cell culture method is carried out in a G-Rex device.