Figure 2
Clonogenic growth and in vivo repopulation of CML SCs. (A) Left: Sorted (>98% pure) CD34+/CD38−/CD26− SC, CD34+/CD38−/CD26+ SC, and CD34+/CD38+ sorted progenitor cells (CP CML) were plated in methylcellulose and incubated at 37°C for 16 days. Then CFU-GM–derived colonies were counted under an inverted microscope (black bars). Thereafter, colonies were picked and BCR/ABL1 mRNA expression was determined by qPCR (white bars). Results represent the mean ± SD of triplicates. Right: Purified CD34+/CD38−/CD26−, CD34+/CD38−/CD26+, and CD34+/CD38+ cells were incubated on irradiated M2-10B4 feeder cells for 3 weeks. Then cells were harvested and cultured in a CFU-GM assay. CFU-GM colonies (black bars) were counted by microscopy and BCR/ABL1 mRNA levels were determined by qPCR (white bars). (B) Experimental setup of xenograft experiments with purified CML cells. First, CML cells (CP) were depleted from lineage+ cells and separated into CD26+ cells and CD26− cells by cell sorting (purity >98%). The 2 fractions were separately injected intravenously into irradiated NSG mice (0.1-0.8 × 106 CD26+ cells per mouse and 0.5-1.5 × 106 CD26− cells per mouse). (C) After 16 weeks or 28 weeks, flushed BM cells of NSG mice injected with CD26− cells (blue symbols) or CD26+ cells (red symbols) were analyzed for the presence of CD33+ myeloid cells (circles) and CD19+ B cells (triangles) by flow cytometry. (D) Flow cytometry patterns of engrafted cells in 2 representative mice, one injected with CD26− SC, where multilineage BCR/ABL1– engraftment was found (upper panel) and one with CD26+ SC, where engraftment with BCR/ABL1+ myeloid cells was seen (lower subpanel). The table shows a summary of engraftment results with percentages of human CD45+ cells, CD33+ cells, and CD19+ cells, and the detection of BCR/ABL1 by qPCR. n.d., not determined.

Clonogenic growth and in vivo repopulation of CML SCs. (A) Left: Sorted (>98% pure) CD34+/CD38/CD26 SC, CD34+/CD38/CD26+ SC, and CD34+/CD38+ sorted progenitor cells (CP CML) were plated in methylcellulose and incubated at 37°C for 16 days. Then CFU-GM–derived colonies were counted under an inverted microscope (black bars). Thereafter, colonies were picked and BCR/ABL1 mRNA expression was determined by qPCR (white bars). Results represent the mean ± SD of triplicates. Right: Purified CD34+/CD38/CD26, CD34+/CD38/CD26+, and CD34+/CD38+ cells were incubated on irradiated M2-10B4 feeder cells for 3 weeks. Then cells were harvested and cultured in a CFU-GM assay. CFU-GM colonies (black bars) were counted by microscopy and BCR/ABL1 mRNA levels were determined by qPCR (white bars). (B) Experimental setup of xenograft experiments with purified CML cells. First, CML cells (CP) were depleted from lineage+ cells and separated into CD26+ cells and CD26 cells by cell sorting (purity >98%). The 2 fractions were separately injected intravenously into irradiated NSG mice (0.1-0.8 × 106 CD26+ cells per mouse and 0.5-1.5 × 106 CD26 cells per mouse). (C) After 16 weeks or 28 weeks, flushed BM cells of NSG mice injected with CD26 cells (blue symbols) or CD26+ cells (red symbols) were analyzed for the presence of CD33+ myeloid cells (circles) and CD19+ B cells (triangles) by flow cytometry. (D) Flow cytometry patterns of engrafted cells in 2 representative mice, one injected with CD26 SC, where multilineage BCR/ABL1 engraftment was found (upper panel) and one with CD26+ SC, where engraftment with BCR/ABL1+ myeloid cells was seen (lower subpanel). The table shows a summary of engraftment results with percentages of human CD45+ cells, CD33+ cells, and CD19+ cells, and the detection of BCR/ABL1 by qPCR. n.d., not determined.

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