Methods of cell culture for adoptive cell therapy

• US 8,956,860 B2
• Filed: 05/18/2012
• Issued: 02/17/2015
• 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., Rooney, Cliona M., Leen, Ann M., Wilson, John R.
• Primary Examiner(s): HOBBS, MICHAEL L
• Application Number:
  US13/475,700
• Publication Number:
  US 20130102075A1
• Time in Patent Office: 1,005 Days
• Field of Search: 435/297.1, 435/373, 435/291.4
• US Class Current: 435/297.1
• 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 gas permeable cell culture and cell recovery device for static cell culture comprising:

an internal volume bounded at least in part by a growth surface and an opposing upper confine, said growth surface comprised of liquid impermeable gas permeable material, at least a portion of said gas permeable material in contact with ambient gas,

a vent,

a medium removal conduit including a medium removal opening,

a cell removal conduit including a cell removal opening,

and when the device is oriented in the static culture position the growth surface is in a horizontal position and is located at least in part below said opposing upper confine, the distance from said growth surface to said cell removal opening is less than the distance from said growth surface to said medium removal opening, and the maximum distance from the upper confine to the growth surface is the maximum potential medium height of the internal volume.

2. The device of claim 1 wherein said maximum potential medium height is beyond 2.0 cm.

3. The device of claim 1 wherein when said device is in the static cell culture position the distance between said growth surface and said medium removal opening is 50% or less than the distance between said growth surface and said upper confine.

4. The device of claim 1 wherein when said device is in the static cell culture position the distance between said growth surface and said medium removal opening is at least 0.2 cm.

5. The device of claim 4 wherein the distance between said growth surface and said medium removal opening is not beyond 2.0 cm.

6. The device of claim 1 wherein when said device is in the static cell culture position said vent resides at a height greater than the height of said medium opening.

7. The device of claim 1 wherein said vent resides at a height beyond the height of said upper confine when the device is in the static cell culture position.

8. The device of claim 1 wherein when said device is in the static cell culture position said cell removal conduit is in contact with said bottom.

9. The device of claim 1 wherein the ratio of the internal volume to the growth surface area is between two and twenty milliliters per square centimeter of growth area.

10. The device of claim 1 including more than one medium removal opening, each medium removal opening residing at a different height from said growth surface when said device is in the static cell culture position.

11. The device of claim 1 including a growth surface support.

12. The device of claim 1 wherein said gas permeable material is comprised of silicone.

13. The device of claim 1 wherein said cell removal opening is oriented at the low point of said internal volume when said device is oriented in a cell removal position.

14. The device of claim 1 wherein said medium removal conduit does not include a filter to keep cells from entering said medium removal conduit.

15. The device of claim 1 including cells and media within the device, the device in a location with ambient gas suitable for cell culture and oriented in the static cell culture position.

16. A method of culturing animal cells with

a gas permeable cell culture and cell recovery device having an internal volume bounded at least in part by a growth surface and an opposing upper confine, said growth surface comprised of liquid impermeable gas permeable material, at least a portion of said gas permeable material in contact with ambient gas,

a vent,

a medium removal conduit including a medium removal opening,

a cell removal conduit including a cell removal opening,

the method comprising:

adding animal cells and media into the device, and culturing cells with said gas permeable cell culture device in a location with ambient gas suitable for cell culture, oriented in a static culture position wherein said growth surface is below said upper confine, said gas permeable material is in a horizontal position, the distance from said growth surface to said cell removal opening is less than the distance from said growth surface to said medium removal opening; and

the maximum distance from the upper confine to the growth surface is the maximum potential medium height of the internal volume.

17. A gas permeable cell culture and cell recovery device for static cell culture comprising:

a cell culture compartment bounded by a growth surface and an opposing upper confine, said growth surface comprised of liquid impermeable gas permeable material, at least a portion of said gas permeable material in contact with ambient gas,

a vent,

a medium removal conduit including a medium removal opening,

a cell removal conduit including a cell removal opening,

and when the device is oriented in the static culture position the growth surface is in a horizontal position and is located at an elevation lower than that of the opposing upper confine, the distance from said growth surface to said cell removal opening is less than the distance from said growth surface to said medium removal opening, and the cell culture compartment is capable of holding media in the cell culture compartment wherein the uppermost location of the media is at a height beyond 2.0 cm from the lowermost location of the media.

18. The device of claim 17 including cells and media within the device, the device in a location with ambient gas suitable for cell culture and oriented in the static cell culture position.

19. A method of culturing animal cells with

a gas permeable cell culture and cell recovery device for static cell culture comprising:

a cell culture compartment bounded by a growth surface and an opposing upper confine, said growth surface comprised of liquid impermeable gas permeable material, at least a portion of said gas permeable material in contact with ambient gas,

a vent,

a medium removal conduit including a medium removal opening,

a cell removal conduit including a cell removal opening,

and when the device is oriented in the static culture position the growth surface is in a horizontal position and is located at an elevation lower than that of the opposing upper confine, the distance from said growth surface to said cell removal opening is less than the distance from said growth surface to said medium removal opening, and the cell culture compartment is capable of holding media in the cell culture compartment wherein the uppermost location of the media is at a height beyond 2.0 cm from the lowermost location of the media;

the method comprising adding animal cells and media into the device and orientating said device in the static culture position in a location with ambient gas suitable for cell culture.

20. The method of claim 19 wherein said animal cells are comprised of T cells.

21. The method of claim 20 wherein said animal cells are comprised of feeder and/or antigen presenting cells.