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. 2024 Sep 18;10(38):eadq1476. doi: 10.1126/sciadv.adq1476

Fig. 7. Preconditioning with HSPC EVs or CCR2 ligands improves HSPC homing to the BM.

Fig. 7.

(A) Schematic detailing intrafemoral injection of HSPC EVs containing Cre mRNA into tdTomato (Ai9) mice. (B) Representative flow plot and quantitative analysis of tdTomato fluorescence in BM ECs isolated from EV-injected marrows. (C) Schematic detailing intrafemoral injection of HSPC EVs before systemic cell transplantation. (D) Flow cytometric quantification of HSPC homing (20 hours after transplantation) toward EV versus sham conditioned marrows (n = 7). (E) Quantification of c-Kit+ Sca-1+ lineage− (KSL) progenitors engrafted in EV versus sham conditioned marrows after 4 weeks (n = 5). (F) Relative gene expression analysis of BM ECs isolated from EV versus sham conditioned marrows (n = 7). (G) Schematic detailing intrafemoral injection of EVs for BM cytokine array profiling (n = 4). (H) Heatmap of fold change (FC; log 2) abundance of cytokines in EV versus sham-conditioned BM. Each column corresponds to the FC of EV:sham-injected marrows in an individual mouse. (I) Schematic detailing intrafemoral injection of individual CCR2L prior to cell transplantation. (J) Flow measurement of HSPC homing toward BM conditioned with individual CCR2L (0.1 ng) versus contralateral sham-injected marrows (n = 4 to 8 per cohort). (K) Relative homing of HSPCs following dose escalation (n = 8) of CCL2 conditioning compared to contralateral sham-injected marrows. (L) Subpopulation-specific engraftment in CCL2-conditioned marrows compared to contralateral sham-injected marrows 4 weeks after transplantation (n = 5). Schematics created using BioRender. *P < 0.05, **P < 0.01, and ***P < 0.001.