Survival, cell cycle progression, and lytic activity of splenic NK cells induced by SCF and IL-2
| c-kit Expression . | Culture Conditions3-150 . | Total Cell No3-151 . | Live (%) Cells . | % Apoptotis . | Cycling (%) Cells . | Specific (%) Lysis . | |
|---|---|---|---|---|---|---|---|
| SCF . | IL-2 . | ||||||
| + | + | high | 180 | 95 | 1.7 | 32 | 78 |
| + | − | high | 124 | 90 | 2.0 | 34 | 84 |
| − | − | high | 105 | 90 | 1.4 | 28 | 89 |
| + | + | low | 83 | 71 | 4.2 | 25 | 79 |
| + | − | low | 76 | 23 | 12 | 11 | 78 |
| − | − | low | 81 | 18 | 21 | 5.0 | 34 |
| c-kit Expression . | Culture Conditions3-150 . | Total Cell No3-151 . | Live (%) Cells . | % Apoptotis . | Cycling (%) Cells . | Specific (%) Lysis . | |
|---|---|---|---|---|---|---|---|
| SCF . | IL-2 . | ||||||
| + | + | high | 180 | 95 | 1.7 | 32 | 78 |
| + | − | high | 124 | 90 | 2.0 | 34 | 84 |
| − | − | high | 105 | 90 | 1.4 | 28 | 89 |
| + | + | low | 83 | 71 | 4.2 | 25 | 79 |
| + | − | low | 76 | 23 | 12 | 11 | 78 |
| − | − | low | 81 | 18 | 21 | 5.0 | 34 |
Splenic c-kit+ and c-kit− NK cells were purified by cell sorting from RAG2−/− mice and plated at 104 cells/well. After 6 days of culture, cells were harvested and enumerated. Dead cells were identified by trypan blue staining. A DNA content analysis was performed by flow cytometry of 7AAD staining, which distinguishes apoptotic cells (hypodiploid) from resting and cycling cells. A 51Cr release assay was performed to assess the lytic activity of effectors against YAC-1 targets at 9:1 E:T ratio. Data are mean values from two experiments.
Cytokine concentrations were 100 ng/mL SCF; 100-1000 units/mL IL-2.
The total cell numbers/well (×103) include both live and dead cells.