American Society of Hematology

Figure 4.

$Figure 4. AML cells release EVs that traffic to surrounding cells, altering the tumor microenvironment. (A) AML EVs were isolated using a spin-column and visualized using transmission electron microscopy (×1000). The negative spin-column fraction was used as a control. To determine whether AML EVs traffic to surrounding cells, we cocultured PBMCs in direct contact or in a 0.4-μM Transwell, with MOLM-14 AML cells pretreated with SYTO RNA dye (530 nm). (B) After 6 hours, MDSCs were quantified for AML SYTO RNA dye using flow cytometry. Healthy donor PBMCs were cultured for 3 days with AML EVs in direct contact or in a 0.4-μM Transwell and then quantified for (C) CD11b+/HLA-DR−/CD33+ MDSCs (expressed as a percentage of immature CD11b+/HLA-DR− myeloid cells) and (D) HLA-DR+/CD11c+ myeloid APCs by flow cytometry (n = 3). *P < .05.$

AML cells release EVs that traffic to surrounding cells, altering the tumor microenvironment. (A) AML EVs were isolated using a spin-column and visualized using transmission electron microscopy (×1000). The negative spin-column fraction was used as a control. To determine whether AML EVs traffic to surrounding cells, we cocultured PBMCs in direct contact or in a 0.4-μM Transwell, with MOLM-14 AML cells pretreated with SYTO RNA dye (530 nm). (B) After 6 hours, MDSCs were quantified for AML SYTO RNA dye using flow cytometry. Healthy donor PBMCs were cultured for 3 days with AML EVs in direct contact or in a 0.4-μM Transwell and then quantified for (C) CD11b⁺/HLA-DR⁻/CD33⁺ MDSCs (expressed as a percentage of immature CD11b⁺/HLA-DR⁻ myeloid cells) and (D) HLA-DR⁺/CD11c⁺ myeloid APCs by flow cytometry (n = 3). *P < .05.

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