Comparison of donor cell distribution in ablated and nonablated mice 20 hours after injection.
(A) Donor cell numbers in injected ablated and nonablated mice (injected/noninjected femur and spleen). Both of the femurs (injected and noninjected) and the spleen were harvested 20 hours after injecting 5 × 105 PKH26-labeled Lin− cells into a femur cavity of untreated or lethally irradiated (950 rad) mice. Bone marrow cells were flushed out from the femurs. Spleens were dissected to wash out spleen cells. Cells from each tissue (injected femur, noninjected femur, and spleen) were counted and subjected to FACS analyses on a MoFlo flow cytometer. Donor cell numbers were calculated from the percentages of donor cells and the total cell count of each tissue. (B) FACS analysis of blood samples from ablated and nonablated mice 20 hours after injection of donor cells into the femur cavity. PKH26-labeled donor cells are present in the blood of ablated mice, but not in the blood of nonablated mice. The autofluorescent events were in the diagonal line and not counted. The positive events fall below the autofluorescent line.
