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. 2020 Jan 2;51:102598. doi: 10.1016/j.ebiom.2019.102598

Fig. 4.

Fig. 4

Characteristics of the CD24+ subset isolated from clinical osteosarcoma samples. (a) Compared to corresponding adjacent tissues, the mRNA expression of CD24 was higher in the 30 primary osteosarcoma samples. Independent experiments were repeated three times. Data are presented as the mean ± standard deviation (t-test, ***p < 0.001; **p < 0.01; and *p < 0.05). (b))1 × 105 CD24+ cells and CD24− cells were sorted by Flow cytometry.CD24− cells isolated from osteosarcoma samples (cases #03, #13, and #15) generated fewer and smaller sarcospheres than CD24+ cells after 7 days of serum deprivation and ultra-low adherent cell culture. (c)1 × 106 CD24+ cells and CD24− cells were sorted by Flow cytometry.The mRNA levels of OCT4, SOX2, NANOG, and BMI1 in CD24+ cells from clinical osteosarcoma samples were higher than those in CD24−cells (cases #04, #05, #08, #09, #11, #12, #14, and #17). Independent experiments were repeated three times. Data are presented as the mean ± standard deviation. Error bars represent the standard deviation (SD) from at least three independent experiments (t-test, ***p < 0.001; **p < 0.01; and *p < 0.05). (d and e) (d) CD24+ cells and CD24− cells were sorted by Flow cytometry. Osteosarcoma cells with different gradients were injected into the medullary cavity of the mouse tibia (No. of mouse = 3 per group). CD24+ osteosarcoma cells from samples (cases #31, #33, #34, #36, and #41) showed a higher tumour-forming incidence than CD24− osteosarcoma cells. (e) Corresponding paraffin-embedded tissues (cases #31 and #36) were processed for H&E staining (scale bar = 100 µm; magnified: scale bar = 50 µm) (see Table S8 for relevant details). (f and g) CD24+ cells and CD24− cells were sorted by Flow cytometry. Osteosarcoma cells with different gradients were injected into the tail vein of the mice(No. of mouse = 3 per group). (f) CD24+ osteosarcoma cells from samples (cases #43, #46, #49, #55, and #58) showed a stronger metastatic capacity than CD24− osteosarcoma cells. (g) H&E staining of corresponding paraffin-embedded lung tissues (#46 and #55) from mice (scale bar = 200 µm; magnified: scale bar = 50 µm) (see Table S9 for relevant details).