From: Lymphocyte expansion in bioreactors: upgrading adoptive cell therapy
Author | Protocol | Stirred system | Expansion | Static control | Purity | Functionality | |||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Stimulation strategy | Medium | Key Supplements | Vessel | Stir. speed [rpm] | Aerat. rate | DO | Seed density [cells/mL] | Volume [ml] | Fold | Days | Fold | Days | Â | Â | |
T cells | |||||||||||||||
 Ou −2019 [90] | IL-2;anti CD3/CD28 beads or mABs | OpTmizer CTS | Serum-free | 2 L Stirred vessel | 70 | 0.01 vvm | 70% | 5 × 105 | 800 | 132–1011 | 4 | – | – | > 99% CD4+ and CD8+ T cells | High CD3 expression (91.7–99.4%); T cell co-stimulatory signaling receptor (ICOS/CD278) elevated; upregulated expression of PD-1/CD279; production of IFNγ decreased by ~ 50% |
 Costariol – 2019 [89] | IL-2; anti CD3/CD28 beads | RPMI 1640 | 10% FBS; Glutamine | Spinner | 35 | – | – | 5 × 105 | 100 | No growth | – | – | – | CD4:CD8 ratio decreased from 4:1 to 1:1 | – |
ambr 250 | 100–200 | 14.25 mL/min 21% O2 | 60% | 5 × 105 | 250 | 23.2 ± 1.3 | 7 | 15.2 ± 3.1 | 7 |  | Effector memory cells increased from 35.69 ± 10.98% to ~ 80%; no significant difference in T cell subpopulation profiles between static and ambr® 250 | ||||
 Ramsborg – 2004 [142] | IL-2; anti CD3/CD28 mABs | AIM V | 2% AP | Spinner | 60 | – | – | 1.5 × 105–3.0 × 105 | 100 | 5.4 (0.5–87) | 15 |  |  | CD3 cells > 90%; CD8 cells preferentially expanded over CD4 cells | – |
 Foster – 2004 [143] | PHA or OKT3; 50 IL2; | RPMI 1640 | 10% AB; Glutamine, HEPES, pyruvate | Spinner | 50 | – | – | 5 × 105 | 100 | > 10 | 7 | > 10 | 7 | NLV–tetramer + CD8+ phenotype > 95% | CD25 decreased exponentially. This behavior did not vary significantly between suspension and static cultures;CTL maintained their specificity toward CMVpp65 |
 Bohnenkamp – 2002 [140] | IL-2; pre anti CD3/CD28 mABs | aMEM | 10% FBS | Spinner | – | – | – | – | – | 394 | 9 | – | – | > 94% CD3+ cells; CD4:CD8 from 2.4:1 to 1:5 (stirred vessel), 1:2.7 (suspension bioreactor) and 1:4.8 (T-flask) | – |
1 L stirred vessel | – | – | – | 1.35 × 105 | – | 44 | 10 | Not different | – | IL-2R was downregulated earlier than in the static T-flask culture; IFNγ secretion assay against a hCMV protein maintained | |||||
Stirred 50–550 mL | – | – | – | 5 × 105 | – | 60 | 10 | Not different | – | – | |||||
 Hilbert −2001 [144] | IL-2;anti CD3/CD28 mABs | RPMI 1640 OR AIMV | 10% FBS; Glutamine | 18 L stirred vessel | – | – | – | – | 18,000 | 11 | 10 | No difference |  | > 90% T-cells | – |
550 mL cell retention reactor | – |  | 20–50% | 3.8 × 105 | 550 | 67 | 5 | – | – | – | – | ||||
 Carswell – 2000 [145] | IL-2, pre PHA or antiCD3 | RPMI 1640 | 10% FBS; Glutamine; pyruvate, neAAs | Spinner | 60 | – | 5% | – | 100 | 1214 ± 374 | 16 | – | – | – | – |
NK cells | |||||||||||||||
 Pierson – 1996 [81] | IL-2; pre anti-CD5/CD8 mABs | (DMEM)/Ham’s F12 | 10% FBS; ascorbic acid; selenite | Reactor | 60 | – | 40% | 106 | 530 | 352 | 33 | – | – | > 90% | NK-specific cytolytic function maintained |
Spinner | 60 | – | – | 106 | 250 | 107 ± 17 | 28 | 43 ± 11 | 28 | 86 ± 9,5% CD56+/CD3-; similar to static | 75% specific lysis of K562; similar to static |