Long constitution capacity of purified HSCs from ossicle bone marrow. HSCs were identified in ossicle bone marrow by immunofluorescent staining with a combination of CD150, CD48, CD41, MECA32 antibodies. The white arrow depicts a HSC that is CD150⁺ (red; A) but negative for CD48, CD41, Lin (green; B), and MECA32 (pink; C). The nuclei are stained with DAPI (blue). (D) HSCs from ossicles were isolated from the combined gate of CD150⁺CD41⁻CD48⁻ and were confirmed to be positive for Sca-1 and c-kit. (E) Five HSCs harvested from ossicles supported long-term multilineage reconstitution in 2 of 4 lethally irradiated mice. Each line represents the frequency of donor-derived myeloid, B, or T cells in a single mouse at 4, 8, 12, and 16 weeks after transplantation. The black line at 0.3% represents the background threshold, meaning that reconstitution is not considered in mice falling below this line.