(A) Experimental schema. HSCs and LSCs were cultured with FACS-sorted BM subsets from normal (CD45.2+) and CML (CD45.1/45.2+) mice for 72 h, transplanted into irradiated WT recipients (CD45.2+), and followed for donor chimerism for 16 weeks after transplantation (n = 5–12 biological replicates per group).
(B and C) Long-term donor chimerism at 16 weeks in the PB of mice transplanted with HSCs (B) and LSCs co-cultured with normal or CML mesenchymal subsets (C).
(D and E) Multilineage donor chimerism in PB (D) and overall donor chimerism in BM (E) at 16 weeks of mice transplanted with LSCs co-cultured with normal or CML mesenchymal subsets. HSCs and LSCs were cultured for 72 h without mesenchymal cells (alone) or co-cultured with normal (NL) or CML 6C3+ cells purified from normal or CML mice at a 1:1 ratio, and cell numbers and immunophenotype (CD45, CD150, and CD48) were analyzed.
(F–J) Representative flow cytometry results (F) and absolute number of CD45+ cells (G), LTHSCs (CD150+CD48−) (H), STHSCs (CD150−CD48−) (I), and MPPs (CD48+) (J).
Error bars represent mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.