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Table 9 Protocol features for stirred systems

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

  

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