Three-dimensional bioreactor for T-cell activation and expansion for immunotherapy

1. A method for T-cell expansion comprising:

supplying a 3D bioreactor of biocompatible polymeric material comprising a plurality of voids having a surface area for cell expansion, said plurality of voids having a diameter D, a plurality of pore openings between said voids having a diameter d, such that D>d and wherein: (a) 90% or more of said voids have a selected void volume (V) that does not vary by more than +/−10.0%; and (b) 90% or more of said pore openings between said voids have a value of d that does not vary by more than +/−10.0%;

providing a polydopamine coating on said bioreactor surface area for cell expansion;

providing a tetrameric protein attached to said polydopamine coating;

providing one or more biotinylated antibodies immobilized on said tetrameric protein;

flowing T-cells through said bioreactor having T-cell receptors where said T-cell receptors bind to said one or more biotinylated antibodies and are activated;

exposing the activated T-cells to a perfusion media containing a signaling molecule to promote T-cell expansion.

2. The method of claim 1 wherein said tetrameric protein comprises avidin or streptavidin.

3. The method of claim 1 wherein said biotinylated antibody comprises anti-CD3 antibody.

4. The method of claim 1 wherein said biotinylated antibody comprises anti-CD3 antibody and anti-CD28 antibody.

5. The method of claim 1 wherein said biotinylated antibody comprises anti-CD3 antibody, anti-CD28 antibody, and anti-CD2 antibody.

6. The method of claim 1 wherein said signaling molecule comprises a cytokine signaling molecule.

7. The method of claim 1 wherein said voids have a diameter (D) of 0.4 mm to 100.0 mm.

8. The method of claim 1 wherein said voids have a diameter (D) of 0.4 mm to 25.0 mm.

9. The method of claim 1 wherein said pores have a diameter (d) in the range of 0.2 mm to 10.0 mm.

10. The method of claim 1 wherein 95.0% or more of said voids indicate a void volume (V) that does not vary by more than +/−10.0%.

11. The method of claim 1 wherein 99.0% to 100% of said voids indicate a void volume (V) that does not vary by more than +/−10.0%.

12. The method of claim 1 wherein 95.0% or more of said pore openings between said voids have a value of d that does not vary by more than +/−10.0%.

13. The method of claim 1 wherein 99.0 to 100% or more of said pore openings between said voids have a value of d that does not vary by more than +/−10.0%.

14. The method of claim 1 wherein at least 90.0% of the voids present have 2 pore openings per void.

15. The method of claim 1 wherein at least 90.0% of the voids present have 8 to 12 pore openings per void.

16. The method of claim 1 wherein said voids have an internal concave surface.

17. The method of claim 1 wherein said voids comprise spherical voids.

18. The method of claim 17 wherein said spherical voids have a packing efficiency of greater than 64.0% in a 3D cylindrical space.

19. The method of claim 1 wherein said 3D bioreactor is formed from a material that has a Tensile Modulus of at least 0.01 GPa.

20. The method of claim 1 wherein said 3D bioreactor is formed from a material not susceptible to hydrolysis during cell cultivation such that the amount of hydrolysis does not exceed 5.0% by weight of the material present.

21. The method of claim 1 wherein said bioreactor has a diameter Φ and a height H and the ratio Φ:H is in the range of greater than 1:1 to 100:1.