Fig. 5

Functional analysis of undivided donor HSCs and MPs. a Individual undivided donor HSCs (LSK CD48⁻ CD150⁺) and PreMeg (Lin⁻ Sca-1⁻ Kit⁺ CD41⁺ CD150⁺ CD16/32⁻) cells were sorted 36 h after transplantation and cultivated in liquid culture media supplemented with mSCF, mTPO, mIl3, and hEpo. Cell composition was analyzed after 12 days using May-Grunwald–Giemsa staining. Colonies (n = 31) for HSCs and (n = 25) PreMegs, 3 independent experiments, 15 mice. In all, 82% HSCs generated colonies (>20 cells) and 79% PreMegs generated >3 megakaryocytes. b Reconstitution experiment using Ubc-GFP mice. c Peripheral blood analysis at 3 weeks after secondary transplantation into lethally irradiated recipients. Donor cell contribution to peripheral blood neutrophils (PMNs) CD11b⁺ Gr1⁺, platelets Ter119⁻CD41⁺, and erythrocytes Ter119⁺. Plots and pictures from 2 independent experiments (n = 5).  LSK repopulation is shown in the upper panel and MP in the bottom panel. We checked the mice every 3–4 weeks for a period of 16 weeks after transplantation but did not find any repopulation from MPs. d Quantification of peripheral blood analysis 3 weeks after transplantation from 2 independent experiments, n = 5. LSK is in red, MP is in black. Statistical significance was determined using unpaired Student’s t-test (*P < 0.05). Data are means +/− s.d.