Three-dimensional bioreactor for viral vector production

A method for closed-loop expansion of viral vector producing cells and viral vector production comprising:

• supplying a three-dimensional bioreactor having an inlet and outlet for the flow of perfusion medium and comprising a plurality of voids having a surface and 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%;
• said surface of said three-dimensional bioreactor is exposed to UV/ozone treatment, plasma gas treatment or coating with extracellular matrix proteins;
• supplying a reservoir for a perfusion medium including oxygen and carbon dioxide;
• supplying a perfusion pump in fluid communication with said inlet and outlet of said three-dimensional bioreactor and said reservoir for said perfusion medium;
• seeding said three-dimensional bioreactor with adherent viral vector producing cells that attach to the three-dimensional bioreactor surface;
• pumping and continuously circulating said perfusion medium from said reservoir into said inlet of said three-dimensional bioreactor and out of said outlet of said three-dimensional bioreactor and promoting viral vector cell expansion;
• introducing a transfection medium to said circulating perfusion medium and transfecting said viral vector cells on said three-dimensional bioreactor surface and producing viral vectors;
• and
• collecting said viral vectors after transfection.