Figure 6: HSC that survive post mortem or in anoxia retain robust regenerative potential after transplantation. (a) Histology of normal mouse BM (H&E staining; n=5 mice).

(b, c) Histology of mouse cadaver BM (2 and 4 days post mortem) showing cell compaction, shrinkage of cytoplasm and oedema (H&E staining after decalcification and paraffin embedding). (d) Percentage of viable whole BM cells several days after storage at 4 °C in anoxia (cell viability assessed by trypan blue exclusion; n=5 mice). (e) Blood chimerism (% of GFP+ cells in circulating blood) in mice post-irradiation and BM transplantation with BM extracted from one femur and collected 0, 1, 2, 3 and 4 days post mortem (n>5 mice per condition). Black bars indicate results with a single BMT, grey bars after one serial transplantation. (f) Immunophenotyping of circulating GFP+ cells after serial BMT showing GFP+ B cells (B220 staining), GFP+ T cells (CD5 staining), GFP+ granulocytes (Gr1 staining), and GFP+ monocytes (CD11b Staining). (g) Reconstitution with BM maintained several days in anoxia. Blood chimerism (% of GFP+ cells in circulating blood) in surviving mice post irradiation and BMT with BM cells kept at 4 °C and in anoxia for 0, 1, 2, 3, 4 days (n> 5 mice per condition). Black bars indicate results with a single BMT and grey bars after one serial transplantation. (h) Reconstitution of all white blood cells lineages as assessed by presence of GFP+ B cells (B220 staining), GFP+ T cells (CD5 staining), GFP+ granulocytes (Gr1 staining) and GFP+ myeloid cells (CD11b staining). Figures display average values of all tests ±s.e.m.