Table 3.
Ex vivo cultivation of natural killer (NK) cells with autologous feeder cells.
| Protocol features | Starting material/culture system | NK cell expansion rate | NK cell purity | NK cell phenotype | NK cell function | Setting | Reference |
|---|---|---|---|---|---|---|---|
| Irr. autologous PBMC (depleted for CD3−/CD56+ cells) + IL-2 + IL-15 | PBMC, CD3 depleted, and CD56 enriched in flasks | 16 (14 days) | 97% NK 0.2% T cells |
Upregulated: NKG2D, DNAM-1, NKp30, NKp44, CD158a, and CD158e | Efficient degranulation and lysis of K562 In vitro |
In vitro | (24) |
| Irr. autologous PBMC activated with OK432, FN-CH296 and OKT-3 + IL-2 | PBMC in flasks and bags | 4,720 (21–22 days) | 91% NK ~12% NK-like T and T |
Strong expression of NKG2D and CD16 | Elevated cytotoxicity that is maintained for up to 4 weeks after infusion to patients | Clinical | (114) |
| Irr. autologous PBMC + OKT-3 + IL-2 | PBMC, CD3 depleted, and CD56 enriched in plates | 169 (14 days) | 84% NK | Upregulated: CD16, CD56, NKG2D, NKp30, and NKp44 | Increased cytotoxicity against tumor cell lines in vitro | In vitro | (115) |
| PBMC, CD3 depleted in flasks and bags | 278–1,097 (21–26 days) | 91–98% NK | Most cells express NKG2D, CD16, CD94, NKp46, KIR2DL1, KIR3DL1, and KIR2DL2/3 | Efficient lysis of tumor cell lines in vitro; persistence in patients up to several months; cytotoxic potential is lost in vivo, while ability for ADCC is maintained | Clinical | (116) | |
| PBMC, CD3 depleted in bags | 691 (14 days) | 98% NK 0.06% T cells |
Upregulated: NKG2C, NKp30, NK44, CXCR4, CD25, CD62L, and CD69 | Increased cytotoxicity against tumor cell lines in vitro; antitumor effect and ADCC activity in a leukemia xenograft mouse model; up to 4 days persistence in patients | Preclinical model | (23) | |
| 758 (14 days) | 98% NK 0.4% T cells |
Clinical | (117) | ||||
| Irr. autologous PBMC (depleted for CD3−/CD56+ cells) + OKT-3 + IL-2 | PBMC, CD3 depleted, and CD56 enriched in plates and flasks | 546 (14 days) | 94.9% NK 2.2% T cells |
Upregulated: NKG2D, NKp30, NKp44, tumor necrosis factor-related apoptosis-inducing ligand, and DNAM-1 Downregulated: NKp80 |
Increased cytotoxicity against tumor cell lines in vitro | In vitro | (113) |
| Irr. autologous PBMC + OKT-3 + IL-2 ± IL-15 | PBMC, CD3 depleted, and CD56 enriched in plates and bags | 117/63 in bags (±IL-15) 993 in plates (19 days) |
Bags: 45% NK 0.6% T cells |
Upregulated: NKG2D, NKp44 | High cytotoxicity against K562 and high productivity of IFN-γ | In vitro | (50) |
| Good manufacturing practice killer cell immunoglobulin-like receptor (KIR) sorted NK cells in bags | 160–390 | ~100% NK >0.01% T cells |
Single KIR + NK cells | Anti-leukemic activity against primary acute myeloid leukemia cells in vitro and in vivo | Preclinical model | (50) |