Recruitment of engrafted donor cells postnatally into the blood with cytokines after in utero transplantation in mice

Background. We have previously shown that MHC-mismatched fetal liver cells can durably engraft in 45% of nondefective fetal mice, although male donor cells were found in the blood in only 8% of female recipients. We postulated that adult bone marrow stem cells would engraft similarly to fetal liver cells and that postnatal administration of cytokines would recruit donor cells into the peripheral circulation. Methods. Bone marrow from C57BL/6 adult male mice was injected into allogeneic BALB/c or congenic C57BL/6 fetal recipients that were 11-13 days old. Engraftment was tested by quantitative polymerase chain reaction for the Y chromosome in female recipients (0.0001% sensitivity), Recipients were injected at 1-2 years of age with rat stem cell factor (SCF) and human granulocyte colony-stimulating factor (G-CSF) for 7 days and tested for donor cells in the blood. Results. The overall engraftment rate (in the blood, spleen, or liver) was 44% in both female allogeneic and congenic bone marrow recipients. Of 49 recipients of bone marrow or fetal liver cells with no evidence of donor cells in the blood before injection, 29 (59%) developed circulating donor cells at some time after cytokine injection for up to 2 months. Twenty-seven of 49 animals were negative in the blood, liver, and spleen before cytokine therapy; 14 of 27 (52%) became positive in the blood after stem cell factor/granulocyte colony-stimulating factor injection. Tolerance to donor skin grafts was not altered by the mobilization of donor cells into the circulation. Conclusions. Adult bone marrow durably engrafts in nondefective mice at a rate similar to that previously obtained with fetal liver. Engrafted donor cells can be mobilized with cytokines into the circulation for up to 2 months even in animals with no evidence of donor cells in the blood, liver, or spleen. Based on these results we estimate that 70-75% of hematopoietically nondefective fetal mice engraft with MHC-mismatched fetal liver or adult bone marrow stem cells.