Figure 4.
Effect of AML cells clearance on AML-MSCs. (A) Principal component analysis (PCA) of AML-MSCs (orange, n = 21), h-MSCs (green, n = 6), and R-MSCs (magenta, n = 10). The x-axis represents the first principal component, PC1, which accounts for the largest variance of mRNA expression, and the y-axis, PC2, explains the second largest variance. (B) Venn diagrams of the differentially expressed genes between AML-MSCs, h-MSCs, and R-MSCs. (C) Cell proliferation by ATP in 2D culture was measured in AML-MSCs (n = 11), h-MSCs (n = 6), and R-MSCs (n = 11) at the indicated time points. (D) Cell density of the murine IL-3–dependent 32D cell line cultured in 2D with AML-MSCs (red line, n = 11), in the presence of IL-3 (black dotted line, n = 11), or without IL-3 (blue line, n = 11), compared with 32D cell line cocultured in 2D with h-MSCs (green line, n = 6), or with R-MSCs (magenta line, n = 11). (E) Quantification (%) of PHA-activated CD3+ T cells expressing CD69 and CD25 after 72 hours of coculture in 2D with h-MSCs (n = 15), AML-MSCs (n = 10), and R-MSCs (n = 6) relative to stroma-free conditions (SF, without MSCs, n = 8). (F) Bio-Plex Pro Human Cytokines 37-Plex Array by using the medium of the MSCs cultured in 3D alone (basal secretome level) or with AML blasts. Color-heatmap shows (each row represents the scaled log2 value of cytokine level, where the 0 value represents the average, and ±4 are range boundaries) cytokine level of AML-MSCs (orange bar, n = 15), h-MSCs (green bar, n = 5), and R-MSCs (magenta, n = 10) cultured alone or cocultured with primary AML cells (the gray bar signed secretome measured in cocultures for 7 days with AML blasts, n = 5). (G) t-SNE (with perplexity score of 25) represents MSCs secretome levels of all samples subdivided for being cocultured in 3D with AML blasts (gray dots, n = 27) or at basal level without blasts (brown dots, n = 27). (H) Radar plots represent the cytokines levels secreted in 3D directly attributable to the AML blasts ([(MSCs + AML cells) − MSCs]/AML) relative to AML-MSCs (orange) or h-MSCs (green). Plots represent 3 different fold change ranges: 0 to 1 (left), 0.5 to 2.5 (middle), and 0.5 to 8.5 (right). *P < .05, **P < .01, ***P < .001, ****P < .0001.

Effect of AML cells clearance on AML-MSCs. (A) Principal component analysis (PCA) of AML-MSCs (orange, n = 21), h-MSCs (green, n = 6), and R-MSCs (magenta, n = 10). The x-axis represents the first principal component, PC1, which accounts for the largest variance of mRNA expression, and the y-axis, PC2, explains the second largest variance. (B) Venn diagrams of the differentially expressed genes between AML-MSCs, h-MSCs, and R-MSCs. (C) Cell proliferation by ATP in 2D culture was measured in AML-MSCs (n = 11), h-MSCs (n = 6), and R-MSCs (n = 11) at the indicated time points. (D) Cell density of the murine IL-3–dependent 32D cell line cultured in 2D with AML-MSCs (red line, n = 11), in the presence of IL-3 (black dotted line, n = 11), or without IL-3 (blue line, n = 11), compared with 32D cell line cocultured in 2D with h-MSCs (green line, n = 6), or with R-MSCs (magenta line, n = 11). (E) Quantification (%) of PHA-activated CD3+ T cells expressing CD69 and CD25 after 72 hours of coculture in 2D with h-MSCs (n = 15), AML-MSCs (n = 10), and R-MSCs (n = 6) relative to stroma-free conditions (SF, without MSCs, n = 8). (F) Bio-Plex Pro Human Cytokines 37-Plex Array by using the medium of the MSCs cultured in 3D alone (basal secretome level) or with AML blasts. Color-heatmap shows (each row represents the scaled log2 value of cytokine level, where the 0 value represents the average, and ±4 are range boundaries) cytokine level of AML-MSCs (orange bar, n = 15), h-MSCs (green bar, n = 5), and R-MSCs (magenta, n = 10) cultured alone or cocultured with primary AML cells (the gray bar signed secretome measured in cocultures for 7 days with AML blasts, n = 5). (G) t-SNE (with perplexity score of 25) represents MSCs secretome levels of all samples subdivided for being cocultured in 3D with AML blasts (gray dots, n = 27) or at basal level without blasts (brown dots, n = 27). (H) Radar plots represent the cytokines levels secreted in 3D directly attributable to the AML blasts ([(MSCs + AML cells) − MSCs]/AML) relative to AML-MSCs (orange) or h-MSCs (green). Plots represent 3 different fold change ranges: 0 to 1 (left), 0.5 to 2.5 (middle), and 0.5 to 8.5 (right). *P < .05, **P < .01, ***P < .001, ****P < .0001.

Close Modal
This Feature Is Available To Subscribers Only

or Create an Account

Close Modal
Close Modal