Culture methods and devices for testing
Improved cell culture devices and related methods that overcome the limitations of prior devices and methods, by creating devices that can integrate a variety of novel attributes. These various attributes include the use of gas permeable material and medium volumes that exceed conventional devices as well as compartments that can facilitate the long term study of high density cultures with reduced disruption of the culture environment, the ability to study the migration of items of interest including substances such as chemokine, track the movement of cells, and monitor cell to cell interactions.
1. A method of assessing functional characteristics of T cells comprising:
adding cancer cells into a culture system including a matrix bio scaffold and allowing an engraftment period for cancer cells to engraft into said matrix bio scaffold,
removing the matrix bio scaffold containing engrafted cancer cells from said culture system and placing the matrix bio scaffold into a compartment of a multi-compartment cell culture apparatus comprising:
at least a first compartment, a second compartment, and a third compartment, each compartment separated from adjacent compartments by a common wall, wherein the wall is mated to a bottom of the compartments, and adapted to hold a volume of medium at a medium volume to bottom footprint ratio of at least 2, each compartment including at least one permanently open passage, wherein the permanently open passage is an opening through the common wall, connecting the bottom of adjacent compartments that allows the contents of one compartment to communicate with another, the bottom of the compartments being gas permeable, the bottom of the compartments and the bottom of the permanently open passages being flat in a common horizontal plane, and not including medium mixing equipment, medium perfusion equipment, or gas pumping equipment; and
adding a volume of medium into the multi-compartment cell culture apparatus,
adding T-Cells into a compartment that differs from the compartment that the matrix bio scaffold containing engrafted cancers cells is added, and allowing a period of time for the T cells to follow a chemokine gradient expressed by the cancer cells, and
measuring the presence of the T Cells in the compartment containing the matrix bio scaffold.
2. The method of claim 1 wherein the apparatus includes a removable lid.
3. The method of claim 1 wherein the culture system is Algimatrix™ 3D Culture System 24 well plate.
4. The method of claim 3 wherein the matrix bio scaffold is Algimatrix™ bio scaffold.
5. The method of claim 1 wherein at least one wall of at least one compartment of the cell culture apparatus is tinted in color.
6. The method of claim 1 , wherein the bottom of at least one compartment of the cell culture apparatus is tinted in color.
7. The method of claim 1 wherein the bottom of at least one compartment of the cell culture apparatus is square or rectangular.
8. The method of claim 1 wherein the bottom of the cell culture apparatus is silicone.
9. The method of claim 8 wherein the silicone is between 0.008 to 0.012 inches thick.
10. The method of claim 1 wherein the bottom of the compartments of the cell culture apparatus have a surface area of 12 cm 2 .
11. The method of claim 1 wherein the walls of the cell culture apparatus allow medium to reside at a height of 5 cm directly above the bottom.
12. The method of claim 1 wherein the opening in each compartment is 2 mm×2 mm.
13. The method of claim 1 claim wherein the cancer cells are bioluminescent.
14. The method claim 1 wherein the T cells are CAR-T cells.
15. The method of claim 1 wherein the permanently open passage of each compartment of the cell culture apparatus does not cut a perpendicular path through the common wall.
16. The method of claim 13 including an additional period of time wherein the T cells initiate killing of the cancer cells killing and anti-tumor activity is measured by a decrease in bioluminescence.
17. The method of claim 1 wherein the cancer cells are CAPAN-1 cancer cells.
18. The method of claim 17 wherein the number of CAPAN-1 cells added to the culture system is 1×106.
19. The method of any claim 1 wherein the engraftment period is 24 hours.
20. The method of claim 1 wherein the period of time is 1 day.
21. The method of claim 1 wherein the period of time is 3 days.
22. The method of claim 16 wherein the additional period of time is between 3 and 28 days.
23. The method of claim 1 wherein an additional volume of medium is not added.
24. A method of assessing functional characteristics of T cells comprising:
Adding cancer cells into a matrix bio scaffold that is attached to the bottom of a compartment of a multi-compartment cell culture apparatus comprising: at least a first compartment, a second compartment, and a third compartment, each compartment separated from adjacent compartments by a common wall, wherein the wall is mated to a bottom of the compartments, and adapted to hold a volume of medium at a medium volume to bottom footprint ratio of at least 2, each compartment including at least one permanently open passage, wherein the permanently open passage is an opening through the common wall, connecting the bottom of adjacent compartments that allows the contents of one compartment to communicate with another, the bottom of the compartments being gas permeable, the bottom of the compartments and the bottom of the permanently open passages being flat in a common horizontal plane, and not including medium mixing equipment, medium perfusion equipment, or gas pumping equipment;
adding a volume of medium into the multi-compartment cell culture apparatus, allowing an engraftment period for the cancer cells to engraft into said matrix bio scaffold;
adding T-Cells into a compartment that differs from the compartment that the matrix bio scaffold is attached, allowing a period of time for the T cells to follow a chemokine gradient expressed by the cancer cells; and
measuring the presence of the T Cells in the compartment containing the matrix bio scaffold.
25. The method of claim 24 wherein the cancer cells are bioluminescent.
26. The method of claim 24 wherein the T cells are CAR-T cells.
27. The method of method of claim 24 including an additional period of time wherein the T cells initiate killing of the cancer cells and anti-tumor activity is measured by a decrease in bioluminescence.
28. The method of claim 27 wherein an additional volume of medium is not added.