Effect of Transduced c-kit on Apoptotic CD34+ CB Cells Analyzed by TUNEL Assay and Flow Cytometry
| . | M5g Neo . | M5g hkit Neo . | ||
|---|---|---|---|---|
| Apoptotic Cells (%) . | No. of Experiments . | Apoptotic Cells (%) . | No. of Experiments . | |
| (I) No SLF | ||||
| Medium | 28.6 ± 6 | 4 | 4.4 ± 3† | 3 |
| TNF-α | 44.1 ± 4* | 4 | 8.5 ± 3† | 3 |
| TGF-β1 | 36.2 ± 5* | 4 | 9.5 ± 4† | 3 |
| (II) SLF (50 ng/mL) | ||||
| Medium | 14.8 ± 4 | 7 | 9.3 ± 3† | 3 |
| TNF-α | 20.1 ± 5* | 7 | 7.8 ± 2† | 3 |
| TGF-β1 | 17.2 ± 3* | 7 | 9.4 ± 3† | 3 |
| . | M5g Neo . | M5g hkit Neo . | ||
|---|---|---|---|---|
| Apoptotic Cells (%) . | No. of Experiments . | Apoptotic Cells (%) . | No. of Experiments . | |
| (I) No SLF | ||||
| Medium | 28.6 ± 6 | 4 | 4.4 ± 3† | 3 |
| TNF-α | 44.1 ± 4* | 4 | 8.5 ± 3† | 3 |
| TGF-β1 | 36.2 ± 5* | 4 | 9.5 ± 4† | 3 |
| (II) SLF (50 ng/mL) | ||||
| Medium | 14.8 ± 4 | 7 | 9.3 ± 3† | 3 |
| TNF-α | 20.1 ± 5* | 7 | 7.8 ± 2† | 3 |
| TGF-β1 | 17.2 ± 3* | 7 | 9.4 ± 3† | 3 |
MACS-separated CD34+ cells were prestimulated with GM-CSF (200 U), IL-3 (200 U), Epo (1 U), and IL-6 (10 ng)/mL and transduced with c-kit or mock viruses as described in Materials and Methods. The cells were washed and incubated with GM-CSF, IL-3, and Epo in the absence or presence of SLF at 50 ng/mL for 48 hours. Cells were washed and labeled with TUNEL reaction mixture and analyzed by flow cytometer. Results are expressed as percent apoptotic cells from an average of 3 to 7 separate experiments.
Significant enhancement in apoptosis with TNF-α or TGF-β1,P < .05.
Significant decrease in apoptosis of c-kit–transduced compared with mock virus-transduced cells, P < .05.