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. 2023 Apr 17;4(4):318–335. doi: 10.1158/2643-3230.BCD-22-0167

Figure 5.

Figure 5. AML-iPSC–derived hematopoietic cells mimic the in vivo clonal dynamics of patients. A, Cell counts of HSPCs derived from the indicated AML-iPSC lines at the indicated days of hematopoietic differentiation liquid culture. N-2.12: a normal iPSC line is shown for comparison. Mean of 3 independent differentiation experiments for each line is shown. B, Number of colonies obtained from 5,000 HSPCs derived from the indicated AML-iPSC lines seeded in methylcellulose assays on day 14 of hematopoietic differentiation. Error bars represent mean and SEM of 1–3 independent differentiation experiments. C, Wright–Giemsa staining of representative cytospin preparations of hematopoietic cells derived from the AML-9.9 line on days 37 and 57 days of hematopoietic differentiation liquid culture, showing predominantly cells with immature myeloid progenitor morphology and prominent mitotic figures. Scale bars, 10 μm. D, Levels of human engraftment (hCD45+) in the BM of NSGS mice transplanted with 1 × 106 HSPCs derived from the indicated 3 pairs of AML-iPSC lines. Each pair is derived from one AML patient and represents an earlier and a later clone, as indicated below the plot. n.s.: not significant (unpaired t test). E, Top panel: schematic overview of the experimental design. HSPCs derived from two lines representing an early (AML-24) and late (AML-4.10) clone from the same AML patient—the latter stably expressing GFP—were mixed 1:1 and intravenously injected into NSGS mice. Bottom: Left: flow cytometry assessment pre-transplant confirming equal mixing of the two clones. Right: flow cytometry assessment of 4 independent mice 5 weeks after transplantation.

AML-iPSC–derived hematopoietic cells mimic the in vivo clonal dynamics of patients. A, Cell counts of HSPCs derived from the indicated AML-iPSC lines at the indicated days of hematopoietic differentiation liquid culture. N-2.12: a normal iPSC line is shown for comparison. Mean of 3 independent differentiation experiments for each line is shown. B, Number of colonies obtained from 5,000 HSPCs derived from the indicated AML-iPSC lines seeded in methylcellulose assays on day 14 of hematopoietic differentiation. Error bars represent mean and SEM of 1–3 independent differentiation experiments. C, Wright–Giemsa staining of representative cytospin preparations of hematopoietic cells derived from the AML-9.9 line on days 37 and 57 days of hematopoietic differentiation liquid culture, showing predominantly cells with immature myeloid progenitor morphology and prominent mitotic figures. Scale bars, 10 μm. D, Levels of human engraftment (hCD45+) in the BM of NSGS mice transplanted with 1 × 106 HSPCs derived from the indicated 3 pairs of AML-iPSC lines. Each pair is derived from one AML patient and represents an earlier and a later clone, as indicated below the plot. n.s.: not significant (unpaired t test). E, Top panel: schematic overview of the experimental design. HSPCs derived from two lines representing an early (AML-24) and late (AML-4.10) clone from the same AML patient—the latter stably expressing GFP—were mixed 1:1 and intravenously injected into NSGS mice. Bottom: Left: flow cytometry assessment pre-transplant confirming equal mixing of the two clones. Right: flow cytometry assessment of 4 independent mice 5 weeks after transplantation.