Table 2.
In vitro models utilizing co-culture of CLL cells with engineered supportive cell lines and soluble factors.
| Stimulatory Cells | Stimulatory Soluble Factors (Concentration) |
Duration in Days |
Purification | CLL Viability | CLL Proliferation | Ref. |
|---|---|---|---|---|---|---|
| irradiated mouse CDw32 L cells |
anti-CD40 mAb (0.5 µg/mL), IL4 (500 U/mL) |
10 | CLL | [3H]TdR uptake 10× higher (vs. control without IL4); increased cell count (until day 8) |
[79] | |
| CDw32 L cells | anti-CD40 moAb (0.5 µg/mL), IL2, IL10 |
3 (up to 9) | CLL | ~80% (~50% *) at day 9 |
weak [3H]TdR uptake compared to healthy B cells | [35] |
| non-irradiated 3T40L cell line | a confluent layer of 3T40L | 3 | ? | 89% (71% *) | - | [21] |
| irradiated 3T40L cell line | 1:17 (3T40L:B cells) | 3 | CLL | 77% (32% *) | - | [80] |
| 3T40L cell line | - | 1–3 | PBMCs/ CLL |
upregulation of genes promoting CLL-cell survival and cell-cycle arrest | [48] | |
| irradiated 3T40L cell line | IL4 (20 ng/mL) | 6 | CLL | 35% (55% *) | cell cycle (6% cells in S-phase vs. 3% in control); [3H]TdR uptake | [42] |
| allogenic human bone marrow stromal cells |
CD40L (1 μg/mL), IL2 (100 U/mL), IL10 (10 ng/mL) |
3.5 | CLL | 35–75% (10–30% *) |
BrdU cell cycle (proliferation induced in 8 of 21 samples); ~4% of cells in S+G2/M phase (DAPI staining) | [61] |
| irradiated 3T40L cell line | CpG (1.5 μg/mL) | 3 | PBMCs |
75% (IGHV unmut.), 70% (IGHV mutated) |
CSFE (proliferation in 80% IGHV unmutated CLL, 25% IGHV mutated CLL at day 5) |
[60] |
| mouse fibroblasts expressing CD31 | IL4 (5 ng/mL) | 7 | PBMCs | 60% (30% *) | CSFE; 4–10% Ki67+ cells | [59] |
| mouse fibroblasts expressing CD40L | IL4 (5 ng/mL) | 7 | PBMCs | 50% (30% *) | CSFE; 8–14% Ki67+ cells | [59] |
| 3T40L cell line | IL21 (25 ng/mL) | 5 | CLL | ~80% (~30% *) at day 8 |
CSFE (~50% divided cells, ~20% w/o IL21); Ki67 | [38] |
| non-irradiated human-bone-marrow stromal (BMSC) cell line UE6E7T-2 | CD40L (1 μg/mL), CpG (1.5 μg/mL), 1:100 (BMSC:B cells) |
3 | PBMCs | 25% (48% *); 32% (60% *) |
↑ S phase; 11% Ki67+ cells (1.18%); ↑ S phase, 7.68% Ki67+ cells (0.81%) |
[34,81] |
| irradiated mouse L cells expressing CD40L |
IL21 (12.5 ng/mL) | 9–10 | CLL | 94% at day 5 | CSFE (proliferation of 49% cells; 18% in control with IL4 instead of IL21) |
[67] |
| non-irradiated human BMSC cell line UE6E7T-2 |
CD40L (1 μg/mL), CpG (1.5 μg/mL), anti-IgM (10 μg/mL) |
2 | PMBCs | 137% (100% *) | 7% Ki67+ cells | [82] |
| 3T40L cell line | IL21 (25 ng/mL) | 5 | PBMCs | as control (~90% *) | CSFE (increased division ~10× compared to control) | [78] |
| HS5 cell line | IL2 (50 ng/mL), CpG (1 µg/mL) | 4 | PBMCs | 70% (82% *) | CSFE (proliferation of 30% cells) | [62] |
| BMF cell line # | CpG (2 µg/mL), IL15 (10 ng/mL) | 7 | CLL | - | CSFE (several generations); Ki67 | [63] |
* Results from control CLL samples cultured at standard conditions are in brackets. # Generated from a long-term culture of bone marrow cells from a CLL patient. ↑ means an increase in percentage of cells in the given cell cycle phase