Determination of functional stem cells by limiting dilution assay
| Genotype /dose, no. of cells . | Positive recipients . |
|---|---|
| wt | |
| 200 000 | 3/3 |
| 100 000 | 3/3 |
| 20 000 | 1/3 |
| 10 000 | 0/3 |
| 5000 | 0/3 |
| Gfi1bko | |
| 200 000 | 3/3 |
| 100 000 | 3/3 |
| 20 000 | 3/3 |
| 5000 | 1/3 |
| Genotype /dose, no. of cells . | Positive recipients . |
|---|---|
| wt | |
| 200 000 | 3/3 |
| 100 000 | 3/3 |
| 20 000 | 1/3 |
| 10 000 | 0/3 |
| 5000 | 0/3 |
| Gfi1bko | |
| 200 000 | 3/3 |
| 100 000 | 3/3 |
| 20 000 | 3/3 |
| 5000 | 1/3 |
The number of functional stem cells was determined in vivo by limiting dilution. Indicated number of pIpC-treated Gfi1bfl/fl and MxCre tg Gfi1bfl/fl (Gfi1bko/ko) (both CD45.2+) bone marrow cells were transplanted with 200 000 CD45.1+ bone marrow cells into lethally irradiated CD45.1+ mice. Approximately 18 weeks after transplantation, peripheral blood was examined for presence of CD45.2+ cells. A percentage higher than 1% was a positive call.