Influence of Cytokines on Numbers of Clonogenic Mammary Carcinoma Cells Detected in Methylcellulose Cultures
| Patient No. . | No. of Colonies per 10,000 Cells (mean ± SD) . | No. of Clonogenic Cells per Culture (mean ± SD) . | ||||||||
|---|---|---|---|---|---|---|---|---|---|---|
| Day 0 . | Day 7 . | Day 0 . | Day 7 . | |||||||
| Controls . | S136E . | TGF-β1 . | S136E + TGF-β1 . | Controls . | S136E . | TGF-β1 . | S136E + TGF-β1 . | |||
| 1 | 68 ± 12 | 83 | 102 | 30 | 21 | 136 ± 12 | 914 | 1172 | 53 | 20 |
| 2 | 3.5 ± 0.5 | 2.5 ± 0.5 | 8 ± 1 | 0 | 0 | 7 ± 0.5 | 33 ± 3.5 | 90 ± 8 | <0.85† | <0.55† |
| 3 | 14 ± 2 | 19 ± 2 | 22 ± 2 | 5 ± 3 | ND | 28 ± 2 | 104 ± 3 | 180 ± 15 | 6.8 ± 0.6 | ND |
| 4 | 0 ± 0 | ND | ND | ND | ND | ND | ND | ND | ND | ND |
| 5 | 0 ± 0 | ND | ND | ND | ND | ND | ND | ND | ND | ND |
| NS | NS | |||||||||
| * | * | |||||||||
| * | * | |||||||||
| Patient No. . | No. of Colonies per 10,000 Cells (mean ± SD) . | No. of Clonogenic Cells per Culture (mean ± SD) . | ||||||||
|---|---|---|---|---|---|---|---|---|---|---|
| Day 0 . | Day 7 . | Day 0 . | Day 7 . | |||||||
| Controls . | S136E . | TGF-β1 . | S136E + TGF-β1 . | Controls . | S136E . | TGF-β1 . | S136E + TGF-β1 . | |||
| 1 | 68 ± 12 | 83 | 102 | 30 | 21 | 136 ± 12 | 914 | 1172 | 53 | 20 |
| 2 | 3.5 ± 0.5 | 2.5 ± 0.5 | 8 ± 1 | 0 | 0 | 7 ± 0.5 | 33 ± 3.5 | 90 ± 8 | <0.85† | <0.55† |
| 3 | 14 ± 2 | 19 ± 2 | 22 ± 2 | 5 ± 3 | ND | 28 ± 2 | 104 ± 3 | 180 ± 15 | 6.8 ± 0.6 | ND |
| 4 | 0 ± 0 | ND | ND | ND | ND | ND | ND | ND | ND | ND |
| 5 | 0 ± 0 | ND | ND | ND | ND | ND | ND | ND | ND | ND |
| NS | NS | |||||||||
| * | * | |||||||||
| * | * | |||||||||
Primary mammary carcinoma cells from the cultures described in Fig2 were plated into semisolid tumor colony assays before (day 0) and after (day 7) treatment with the indicated cytokines. Colonies were scored on day 17 to 20 after plating and absolute numbers of clonogenic tumor cells per culture were calculated by multiplying the clonogenicity of tumor cells with the number of total cells in the cultures. The tumor cell origin of colonies was confirmed in randomly picked colonies by immunostaining with FITC-anticytokeratin MoAbs. Results represent mean values ± SD of duplicate determinations. Statistically insignificant (NS, P > .05) or significant (*P < .05) differences between treated and control groups were obtained using Student’s t-test after normalizing data of patients no. 1 through 3.
Abbreviations: S136E, 10 ng/mL SCF, 3 ng/mL IL-1β, 100 ng/mL IL-3, 100 ng/mL IL-6, 1 U/mL EPO; TGF-β1, 30 ng/mL TGF-β1; ND, not done.
Detection limit.