METHODS OF CELL CULTURE FOR ADOPTIVE CELL THERAPY

• US 20150175966A1
• Filed: 12/22/2014
• Published: 06/25/2015
• Est. Priority Date: 12/08/2009
• Status: Active Grant

• Current Assignee: Baylor College of Medicine (Baylor College), Wilson Wolf Manufacturing Corporation
• Original Assignee: Baylor College of Medicine (Baylor College)
• Inventors: Vera, Juan F., Rooney, Cliona M., Leen, Ann M., Wilson, John R.
• Granted Patent: US 9,567,565 B2
• Time in Patent Office: Days
• US Class Current: 1/1
• CPC Class Codes: A61K 2039/5156 expressing foreign proteins
• CPC Class Codes: A61K 2039/5158 Antigen-pulsed cells, e.g. T-cells
• CPC Class Codes: A61K 39/0011 Cancer antigens
• CPC Class Codes: C07K 14/7051 T-cell receptor (TcR)-CD3 complex
• CPC Class Codes: C07K 2319/03 containing a transmembrane segment
• CPC Class Codes: C12M 23/24 Gas permeable parts
• CPC Class Codes: C12N 2501/2302 Interleukin-2 (IL-2)
• CPC Class Codes: C12N 2502/11 blood or immune system cells
• CPC Class Codes: C12N 5/0638 Cytotoxic T lymphocytes [CTL] or lymphokine activated killer cells [LAK]

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. A static culture method of expanding a population of cells with a gas permeable cell culture device having

a growth surface comprised of liquid impermeable gas permeable material, at least a portion of said gas permeable material in contact with ambient gas,

the static culture method comprising:

adding cells and medium into the gas permeable cell culture device, the ratio of the volume of said medium to the surface area of said growth surface being 3 ml/cm² to 20 ml/cm², orienting the gas permeable cell culture device in a position wherein ambient gas suitable for cell culture makes contact with said gas permeable material and cells reside upon at least a portion of said gas permeable material, and

allowing static cell culture for a period of time and not placing additional medium into the gas permeable cell culture device during said period of time, and

said period of time is at least 3 days, and

adding at least one type of cytokine after the onset of static cell culture.

2. The method of claim 1 wherein said period of time is at least 4 days.

3. The method of claim 1 wherein said period of time is at least 5 days.

4. The method of claim 1 wherein said period of time is at least 6 days.

5. The method of claim 1 wherein said period of time is at least 7 days.

6. The method of claim 1 wherein said period of time is at least 8 days.

7. The method of claim 1 wherein said period of time is at least 9 days.

8. The method of claim 1 wherein said period of time is at least 10 days.

9. The method of claim 1 wherein said period of time is at least 11 days.

10. The method of claim 1 wherein said period of time is at least 12 days.

11. The method of claim 1 wherein said period of time is at least 13 days.

12. The method of claim 1 wherein said period of time is at least 14 days.

13. The method of claim 1 wherein the ratio of the quantity of cells to the surface area of said growth surface at the onset of static cell culture is at least 2.0×

10⁶ cells/cm².

14. A static culture method of expanding a population of cells with a gas permeable cell culture device having

a growth surface comprised of liquid impermeable gas permeable material, at least a portion of said gas permeable material in contact with ambient gas,

the static culture method comprising:

adding cells and medium into the gas permeable cell culture device, the ratio of the volume of said medium to the surface area of said growth surface being 3 ml/cm² to 20 ml/cm², and

orienting the gas permeable cell culture device in a position wherein ambient gas suitable for cell culture makes contact with said gas permeable material and cells reside upon at least a portion of said gas permeable material, and

allowing static cell culture over a period of time without placing additional medium into the gas permeable cell culture device, and

during said period of time the ratio of the quantity of cells to the surface area of said growth surface becomes at least 4×

10⁶ cells/cm².

15. The method of claim 14 wherein said ratio of the quantity of cells to the surface area of said growth surface becomes at least 4.5×

10⁶ cells/cm².

16. The method of claim 14 wherein said ratio of the quantity of cells to the surface area of said growth surface becomes at least 5.0×

10⁶ cells/cm².

17. The method of claim 14 wherein said ratio of the quantity of cells to the surface area of said growth surface becomes at least 5.7×

10⁶ cells/cm².

18. The method of claim 14 wherein said ratio of the quantity of cells to the surface area of said growth surface becomes at least 7×

10⁶ cells/cm².

19. The method of claim 14 wherein said ratio of the quantity of cells to the surface area of said growth surface becomes at least 7.5×

10⁶ cells/cm².

20. The method of claim 14 wherein said ratio of the quantity of cells to the surface area of said growth surface becomes at least 8.1×

10⁶ cells/cm².

21. The method of claim 14 wherein said ratio of the quantity of cells to the surface area of said growth surface becomes at least 8.5×

10⁶ cells/cm².

22. The method of claim 14 wherein said ratio of the quantity of cells to the surface area of said growth surface becomes at least 10×

10⁶ cells/cm².

23. The method of claim 14 wherein said ratio of the quantity of cells to the surface area of said growth surface becomes at least 11.0×

10⁶ cells/cm².