NGFR Transgene Expression in CFC-Derived Progeny
| Cells . | Vectors . | CFC Frequency (%) . | % of Individual Colonies That Show Transgene Presence (no.+/no. analyzed) . | % of Pooled Cells That Show Transgene Expression at Day 14 . |
|---|---|---|---|---|
| MINGFR | 26.3 ± 0.9 | 28 (16/64) | 12.5 | |
| (81% CD34+Lin−, 34% NGFR+) | ||||
| Unsorted | LINGFR | 25.1 ± 0.2 | 34 (22/65) | 7.7 |
| (81% CD34+Lin−, 32% NGFR+) | ||||
| None | 23.6 ± 1.2 | ND | 0.5% | |
| (81% CD34+Lin−) | (background) | |||
| Sorted | MINGFR | 22.9 ± 3.5 | ND | 78.0 |
| (NGFR+CD34+Lin−) | LINGFR | 19.9 ± 0.9 | ND | 52.6 |
| Cells . | Vectors . | CFC Frequency (%) . | % of Individual Colonies That Show Transgene Presence (no.+/no. analyzed) . | % of Pooled Cells That Show Transgene Expression at Day 14 . |
|---|---|---|---|---|
| MINGFR | 26.3 ± 0.9 | 28 (16/64) | 12.5 | |
| (81% CD34+Lin−, 34% NGFR+) | ||||
| Unsorted | LINGFR | 25.1 ± 0.2 | 34 (22/65) | 7.7 |
| (81% CD34+Lin−, 32% NGFR+) | ||||
| None | 23.6 ± 1.2 | ND | 0.5% | |
| (81% CD34+Lin−) | (background) | |||
| Sorted | MINGFR | 22.9 ± 3.5 | ND | 78.0 |
| (NGFR+CD34+Lin−) | LINGFR | 19.9 ± 0.9 | ND | 52.6 |
Two days after transduction using mPB CD34+Lin− cells, unsorted and the transgene-expressing subset of CD34+Lin−cells were each plated into methylcellulose culture. Colonies were enumerated at day 14. The presence of the retroviral DNA in individually picked colonies was determined by PCR, and transgene expression in CFC-derived cells was determined by flow cytometry of pooled colonies.
Abbreviation: ND, not done.