Lin− HSC transduction
| Vector . | Transduction conditions no. animals positive/no. animals tested . | |
|---|---|---|
| 24 hours without cytokines* . | 96 hours with cytokines† . | |
| Group 1 | ||
| Eco-MLV | 0/15 | 12/15 |
| M-GFP | 1/15 | 10/15 |
| Group 2 | ||
| Eco-MLV | 0/15 | 14/15 |
| L-GFP | 1/15 | 10/15 |
| Vector . | Transduction conditions no. animals positive/no. animals tested . | |
|---|---|---|
| 24 hours without cytokines* . | 96 hours with cytokines† . | |
| Group 1 | ||
| Eco-MLV | 0/15 | 12/15 |
| M-GFP | 1/15 | 10/15 |
| Group 2 | ||
| Eco-MLV | 0/15 | 14/15 |
| L-GFP | 1/15 | 10/15 |
HSC indicates hematopoietic stem cell; MLV, Moloney murine leukemia virus;GFP, green fluorescent protein.
Animals were tested by polymerase chain reaction analysis of peripheral blood DNA 13 to 16 weeks after transplant for presence of the proviruses. Animals from group 1 were transplanted with cells transduced with Eco-MLV-LacZ and M-GFP, animals from group 2 were transplanted with cells transduced with Eco-MLV-LacZ and L-GFP. The results from 3 different experiments are combined.
Cells were exposed to the vectors for 24 hours with medium changed after 12 hours in Dulbecco modified Eagle medium (DMEM) and 15% fetal calf serum (FCS) without cytokines.
Cells were exposed to the vectors for 96 hours with medium changed every 24 hours. The medium contained DMEM-15% FCS and interleukin 3 (IL-3), IL-6, and stem cell factor.