Figure 1.

Generation of HPCs from hiPSCs. (A) An outline of the protocol is shown. During the first 12 days, HPCs were generated from hiPSC using the StemDiff™ Hematopoietic kit, and were subsequently differentiated to mature NK cells on OP9-DL1 feeder cells over three weeks. (B) The morphology of EBs and emerging HPCs during 12 days of differentiation is shown. On day -1 a confluent iPSC layer has been cut into uniform squares. Day 0: EBs of different sizes occur. Day 3 and 5: EBs start to enlarge and attach to the culture vessel surface. Subsequently, attached EBs continue to spread forming large clumps. From day 7, progenitor cells appear as bright and round cells budding off from the attached structures. The bar represents 1mm and 100µm, corresponding to 4x and 20x magnification, respectively. (C) Representative flow cytometric analysis of CD34, CD43 and CD45 expression on day 12. (D) Percentages of CD34⁺, CD43⁺, CD45⁺ and CD45⁺CD34⁺ cells obtained from multiple independent flow cytometric experiments on day 12 of HPC differentiation (n = 5, CD43: n=4). Bars represent mean and standard deviation.