Reconstitution analysis in lethally irradiated mice transplanted with cultured hematopoietic stem cells on ALCAM-transfected stromal cells
| Primary fetal liver stem cells . | Cultured cells . | Total % Ly5.1 cells with vector-transfected cells . | Total % Ly5.1 cells with ALCAM-transfected cells (GM2) . | ||
|---|---|---|---|---|---|
| 7 days . | 25 days . | 7 days . | 25 days . | ||
| 10.1% ± 5.8% (500 cells/mouse) | One sixth of culture | ND | 0.7% ± 0.2% | ND | 3.2% ± 2.1% |
| 18.2% ± 14.3% (1000 cells/mouse) | One third of culture | 1.5% ± 1.2% | 0.9% ± 0.2% | 9.4% ± 4.4% | 6.2% ± 0.9% |
| Primary fetal liver stem cells . | Cultured cells . | Total % Ly5.1 cells with vector-transfected cells . | Total % Ly5.1 cells with ALCAM-transfected cells (GM2) . | ||
|---|---|---|---|---|---|
| 7 days . | 25 days . | 7 days . | 25 days . | ||
| 10.1% ± 5.8% (500 cells/mouse) | One sixth of culture | ND | 0.7% ± 0.2% | ND | 3.2% ± 2.1% |
| 18.2% ± 14.3% (1000 cells/mouse) | One third of culture | 1.5% ± 1.2% | 0.9% ± 0.2% | 9.4% ± 4.4% | 6.2% ± 0.9% |
Day 14 fetal liver cells (Ly5.1) were purified for Lin−Sca-1+c-Kit+ cells, and 3000 stem cells were cultured with ALCAM- or vector-transfected cell lines for 7 days or 25 days. Cells were harvested, and one third of culture (corresponding to 1000 stem cells) or one sixth of culture (corresponding to 500 stem cells) was transplanted with 1 × 106 Ly5.2 competitor bone marrow cells into lethally irradiated Ly5.2 mice. The expression of the donor-type Ly5.1 marker on peripheral blood cells was examined 12 weeks after transplantation. Three independent experiments were performed, and representative data were obtained from one experiment.
ND indicates not done.