Rapid expansion in the WAVE bioreactor of clinical scale cells for tumor immunotherapy

Abstract

Cell-based immunotherapy using natural killer (NK) cells, cytokine-induced killer (CIK) cells and dendritic cells (DCs) is emerging as a potential novel approach in the auxiliary treatment of a tumor. However, non-standard operation procedure, small-scale cell number, or human error may limit the clinical development of cell-based immunotherapy. To simplify clinical scale NK cells, CIK cells and DCs expansions, we investigated the use of the WAVE bioreactor, a closed system bioreactor that utilizes active perfusion to generate high cell numbers in minimal volumes. We developed an optimized rapid expansion protocol for the WAVE bioreactor that produces clinically relevant number of cells for our adoptive cell transfer clinical protocols. The high proliferative rate, surface phenotypes, and cytotoxicity of these immune cells, as well as the safety of cultivation were analyzed to illuminate the effect of WAVE bioreactor. The results demonstrated that the benefit of utilizing modern WAVE bioreactors in cancer immunotherapy was simple, safe, and flexible production.

Propofol Promotes Activity and Tumor-Killing Ability of Natural Killer Cells in Peripheral Blood of Patients with Colon Cancer

“…Immunotherapy has long been an important topic in cancer research, including adoptive immunotherapy and cytokine-induced killer therapy [ 26 , 27 ]. With the deepening of understanding of immune cells, the number of immunotherapy methods for cancers is growing.…”

Adoptive Immunotherapy beyond CAR T-Cells

“…The bag ports are designed to allow continuous medium supply and circulation through a special filter that is not permeable for the cells. The WAVE ® bioreactor is often used for cultivation of the majority of therapeutic immune cells [ 44 , 100 ]; however, it was shown to be better suited for manufacturing NK-cells than DCs [ 101 ]. Manufacturing of therapeutic macrophages and CAR M-cells can also be performed in suspension with motion-based or stirred tank bioreactors; however, this might require using special low-adhesion materials [ 102 ].…”

Lymphocyte expansion in bioreactors: upgrading adoptive cell therapy

“…Additionally, perfusion enables cells to be expanded above 10 7 cells/mL, supporting high volume cultures to be carried out in a single bag with a significantly reduced volume (some bioreactor cultures need about half the amount of media to harvest 10 10 cells, as compared to static conditions). Since bags are single use there is no need for cleaning validation, they provide a ready-to-use closed system decreasing turnaround time and resource requirements, significantly reducing costs in GMP operations [ 97 ]. Consequently, this platform is frequently used academically and industrially during phase 1 and 2 clinical trials [ 12 ].…”

“… Spanholtz [ 104 ] - 2011 Used a GM-CSF. G-CSF, IL-6, IL-7, IL-15 GBGM 10% HS CD34+ UCB 1.0 × 10 6 – from 250 WAVE Bioreactor System 2/10 and BIOSTATH CultiBag RM 10 0.1–0.2 6 Media addition to adjust cell density Rujkijyanont [ 105 ] - 2013 500 CD56- (APCs); IL15 OKT3 SCGM 5% AB CD56+ from PBMCs 1.0 × 10 6 2–20 from 200 WAVE Bioreactor 5–9 – – Media addition to adjust cell density Lapteva [ 106 ] - 2014 500 K562-41BBL-mbIL-15 (APCs) SCGM 10% FBS CD56+ from PBMCs 2.0 × 10 5 – – WAVE Bioreactor 6 – 6 – Meng [ 97 ] - 2018 Used a Pre OK-432 Xvivo 15 1% AP PBMC – 3 3000 GE Xuri W25 7 – 6 Perfusion (parameters not specified) TILs Sadeghi [ 107 ] - 2011 600 PBMCs (APCs); OKT3 RPMI 5% AB; GLN 12 mM, 25 mM HEPES; BME TIL 5–10.0 × 10 7 (TOTAL) 2 1000 Wave Bioreactor System 2/10 (GE) 10 – 6 Perfusion: 350–1000 mL/day to maintain glucose and GLN in a range of 1.5–2 g/L and ~ 2–4 mM, respectively. Somerville [ 64 ] - 2012 3000 Pre – allogen...…”