Fig. 3

Hematopoiesis analysis after co-culture with Casepase-3^−/− BA-MSCs. A Schematic of the experimental procedures. The co-culture of HSPCs from wild-type (WT) mice (CD45.1) and BA-MSCs from Caspase-3^−/− (KO) mice (CD45.2) was performed in MEM-alpha medium. After 48 h, suspending HSPCs were harvested for flow cytometry and colony-forming assay. Ctrl: HSPCs cultured alone; KO: HSPCs cultured with Caspase-3^−/− BA-MSCs; WT: HSPCs cultured with WT BA-MSCs. In addition, the obtained HSPCs (CD45.1) were mixed with WT helper cells (CD45.2), and injected into 8.0 Gy-irradiated recipient mice (CD45.2) through ophthalmic vein. Engraftment efficiency in recipients was monitored by donor contribution of CD45.1⁺ cells in the peripheral blood and bone marrow. B Flow cytometry analysis of the percentage of LT-HSC, ST-HSC, MPP, CMP and GMP after co-culture (n = 3). C Flow cytometry analysis of the percentage of CD11b⁺/Gr-1⁺ myeloid cells after co-culture (n = 3). D Morphology of colony-forming units of granulocyte–macrophage colonies (CFU-G, CFU-M, and CFU-GM). E Analysis of total colony numbers (including CFU-E, BFU-E, CFU-G, CFU-M, CFU-GM, and CFU-GEMM) at 7 and 14 days for the detection of HSPCs after co-culture (n = 5). F Analysis of myeloid colony numbers (including CFU-G, CFU-M, and CFU-GM) at 14 days for the detection of HSPCs after co-culture (n = 5). G The analysis of peripheral blood cells was performed every four weeks post-transplantation (WT: n = 4; KO: n = 3). H The assessment of progenitor subpopulations in the bone marrow was performed at 16 weeks after mice sacrifice (WT: n = 4; KO: n = 3). All data are presented as mean ± SEM. ns: not significant, *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001