Figure 4.

Acute myeloid leukemia (AML) patient-specific hematopoietic progenitor cells (HPCs) are capable of normal in vitro differentiation to mature blood cells and are devoid of leukemia-associated aberration. (A): Experimental strategy used to characterize patient-specific, putative HPCs in vitro. Methodologies used to assess normal hematopoietic functional capacity are indicated in red. (B): Putative HPC functionality assessed by multilineage differentiation capacity in in vitro colony-forming unit assay. Bars represent mean frequencies of mature hematopoietic colonies generated + SEM (n = 3 independent experiments per patient-specific HPC line). AML patient Fib iPSC-derived HPCs generate all mature lineages, consistent with healthy patient Fib iPSC-derived HPCs. (C): Representative mature hematopoietic colonies derived from patient-specific HPCs. Scale bars represent 100 µm. (D): Representative single-cell morphologies following Giemsa-Wright staining performed on individual hematopoietic colonies (n ≥ 3 colonies analyzed per patient-specific HPC line). Scale bars represent 10 µm. (E): Fluorescence in situ hybridization performed in total mature hematopoietic colonies derived from patient-specific HPCs. Aberration identified in matched patient AML bone marrow was not detected. Adjacent plots depict the number of nuclei (blue circle) scored; 500 nuclei were analyzed to exclude 1% genetic mosaicism with 99% confidence ⁴¹.