Replacement of BM cells in hemophilia A mice after transplantation of 2 × 106 BM cells. (A) Timeline indicating animal groups subjected to BM transplantation (Tx), including no further treatment (NT) or J0-2 antibody, MCT, and CCl4 followed by assays for therapeutic correction. (B) Flow cytometry showing BM chimerism after 2 months in recipients of Rosa 26 BM with LacZ expression detected by FDG. Panels on left are negative controls. R3 gate in panels on right indicates BM repopulating cells. BM chimerism was extensive, including mice with BM Tx and J0-2, MCT, and CCl4. (C) Flow cytometry after 2 months in recipient of GFP+ BM cells showing chimerism of 90% in blood and 60% in BM. (D) Flow cytometry after 2 months in recipients (n = 7) of CD45.1 BM cells showing > 80% chimerism. (E) GFP+ donor-derived cells in liver sinusoids (green, arrow) 6 months after transplant. Stains included DAPI for nuclei (blue) and F4/80 for KC (red). Donor BM-derived KCs appear yellow in merged image (arrow, right panel). Original magnification ×400.
