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. 2018 Mar 15;7(6):468–476. doi: 10.1002/sctm.17-0189

Figure 3.

Figure 3

CD11a and EPCR alone are sufficient to sort a rare population enriched for HSCs. (A): Sorting strategy using only CD11a and EPCR as HSC markers. CFP+ CD11a– EPCR+ and Tomato+ “Not 11a/EPCR” (not CD11a– EPCR+) cells (and vice versa) were sorted and co‐transplanted in a competitive setting at physiological ratios. 850 CD11a– EPCR+ and 500,000 Not 11a/EPCR were transplanted into each recipient. Percentages of cells within each gate are shown. (B, C): Time‐course analysis of total blood (B) and blood granulocyte (C) chimerism from CD11a– EPCR+ and Not 11a/EPCR sources in primary recipients 4, 8, and 16 weeks (W) post‐transplant, and in secondary recipients (separated by vertical dashed line) at W6 following secondary transplant. CFP+ donor‐derived cells are represented by outlined symbols and Tomato+ donor‐derived cells with borderless symbols. Not all primary recipients were selected for secondary transplantation. (D): Donor chimerism of HSPCs in the BM of primary recipients transplanted with “CD11a– EPCR+” and “Not 11a/EPCR” sorted cells 17 weeks post‐transplant. HSPCs are defined as Ter119– CD27+ Sca‐1+ Kit+. CFP+ donor‐derived cells are represented by outlined symbols and Tomato+ donor‐derived cells with borderless symbols. (E): Sorting strategy to compare CD11a– EPCR+ (11a/EPCR) to Ter119– CD27+ Kit+ Sca‐1+ CD34– CD150+ CD48– (SLAMKLS34). 680 CFP‐expressing 11a/EPCR and 60 Tomato‐expressing SLAMKLS34 (and vice versa) were sorted and co‐transplanted in a competitive setting. 250,000 nonlabeled WBM was used as helper for each recipient. (F–H): Time‐course analysis of total blood (F) and blood granulocyte (G) chimerism from 11a/EPCR and SLAMKLS34 sources in primary recipients 4, 9, and 12 weeks (W) post‐transplant and HSPC chimerism (H) at W13. CFP+ donor‐derived cells are represented by outlined symbols and Tomato+ donor‐derived cells with borderless symbols. *, p ≤ .05; **, p ≤ .01; ***, p ≤ .001 (Student's unpaired t test). “Not 11a/EPCR” = not CD11a‐EPCR+. Abbreviations: BM, bone marrow, EPCR, endothelial protein C receptor; HSPC, hematopoietic stem and progenitor cells.