Methods of cell culture for adoptive cell therapy
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/cm 2 if the growth surface is not comprised of gas permeable material and up to 2 ml/cm 2 if the growth surface is comprised of gas permeable material. The desired cells are at a surface density of less than 0.5×10 6 cells/cm 2 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 1.25×10 5 cells/cm 2 .
1. A static culture method for 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 an ambient gas,
the method comprising:
adding a population of desired cells and a population of antigen presenting and/or feeder cells and a medium into the gas permeable cell culture device, a ratio of the volume of said medium to a surface area of said growth surface being 2.5 ml/cm 2 to 15 ml/cm 2 ,
commencing a static cell culture with the ambient gas suitable for the static cell culture making contact with said gas permeable material and in an orientation wherein said desired and antigen presenting and/or feeder cells reside upon at least a portion of said gas permeable material,
not adding fresh medium from the time the static culture commenced,
terminating the static cell culture beyond 4 days from the time the static cell culture commenced, and
when the size of the population of desired cells exceeds the size of the population of antigen presenting and/or feeder cells.
2. The method of claim 1 wherein the static cell culture is not terminated until some point in time beyond 5 days from the time the static cell culture commenced.
3. The method of claim 1 wherein the static cell culture is not terminated until some point in time beyond 6 days from the time the static cell culture commenced.
4. The method of claim 1 wherein the static cell culture is not terminated until some point in time beyond 7 days from the time the static cell culture commenced.
5. The method of claim 1 wherein the static cell culture is not terminated until some point in time beyond 8 days from the time the static cell culture commenced.
6. The method of claim 1 wherein the static cell culture is not terminated until some point in time beyond 9 days from the time the static cell culture commenced.
7. The method of claim 1 wherein the static cell culture is not terminated until some point in time beyond 10 days from the time the static cell culture commenced.
8. The method of claim 1 wherein the static cell culture is not terminated until some point in time beyond 11 days from the time the static cell culture commenced.
9. A static culture method for expanding a population of cells with a gas permeable cell culture device having
a growth surface comprised of a liquid impermeable gas permeable material, at least a portion of said gas permeable material in contact with an ambient gas,
the method comprising:
adding a population of desired cells and a population of antigen presenting and/or feeder cells and a medium into the gas permeable cell culture device, the ratio of the volume of said medium to a surface area of said growth surface being at least 5 ml/cm 2 to 15 ml/cm 2 ,
commencing a static cell culture with the ambient gas suitable for a cell culture making contact with said gas permeable material and in an orientation wherein said desired and of antigen presenting and/or feeder cells reside upon at least a portion of said gas permeable material,
not adding fresh medium from the time the static cell culture commenced,
terminating the static cell culture after 4 days from the time the static cell culture commenced, and
when the size of the population of desired cells exceeds the size of the population of antigen presenting and/or feeder cells.
10. The method of claim 9 wherein the static cell culture is not terminated until some point in time beyond 5 days.
11. The method of claim 9 wherein the static cell culture is not terminated until some point in time beyond 6 days.
12. The method of claim 9 wherein the static cell culture is not terminated until some point in time beyond 7 days.
13. The method of claim 9 wherein the static cell culture is not terminated until some point in time beyond 8 days.
14. The method of claim 9 wherein the static cell culture is not terminated until some point in time beyond 9 days.
15. The method of claim 9 wherein the static cell culture is not terminated until some point in time beyond 10 days.
16. The method of claim 9 wherein the static cell culture is not terminated until some point in time beyond 11 days.
17. A static culture method for expanding a population of cells with a gas permeable cell culture device having
a growth surface comprised of a liquid impermeable gas permeable material, at least a portion of said gas permeable material in contact with an ambient gas,
the method comprising:
adding a population of desired cells and a population of antigen presenting and/or feeder cells and a medium into the gas permeable cell culture device, a ratio of the volume of said medium to a surface area of said growth surface being 10 ml/cm 2 to 15 ml/cm 2 ,
commencing a static cell culture with ambient gas suitable for the static cell culture making contact with said gas permeable material and in an orientation wherein said desired and antigen presenting and/or feeder cells reside upon at least a portion of said gas permeable material,
not adding fresh medium from the time the static culture commenced,
terminating the static cell culture after 4 days from the time the static cell culture commenced, and
when the size of the population of desired cells exceeds the size of the population of antigen presenting and/or feeder cells.
18. The method of claim 17 wherein the static cell culture is not terminated until some point in time beyond 5 days.
19. The method of claim 17 wherein the static cell culture is not terminated until some point in time beyond 6 days.
20. The method of claim 17 wherein the static cell culture is not terminated until some point in time beyond 7 days.
21. The method of claim 17 wherein the static cell culture is not terminated until some point in time beyond 8 days.
22. The method of claim 17 wherein the static cell culture is not terminated until some point in time beyond 9 days.
23. The method of claim 17 wherein the static cell culture is not terminated until some point in time beyond 10 days.
24. The method of claim 17 wherein the static cell culture is not terminated until some point in time beyond 11 days.