Derivation of functional mature neutrophils from human embryonic stem cells. Blood 113: 6584–6592

Human embryonic stem cells (hESC) proliferate infinitely and are pluripotent. Only a few reports, however, describe specific and efficient methods to induce hESC to differentiate into mature blood cells. It is important to determine whether and how these cells, once generated, behave similarly with their counterparts produced in vivo. We developed a method to induce the differentiation of hESC into mature neutrophils. Embryoid bodies were formed with bone morphogenic protein-4, stem cell factor (SCF), Flt-3 ligand (FL), interleukin (IL)-6/IL-6 receptor fusion protein (FP6), and thrombopoietin (TPO). The cells derived from the embryoid bodies were cultured on a layer of irradiated OP9 cells with the combination of SCF, FL, FP6, IL-3, and TPO, which was later changed to granulocyte-colony stimulating factor. Morphologically mature neutrophils were obtained in approximately 2 weeks with a purity and efficiency sufficient for functional analyses. The population dominantly containing mature neutrophils (hESC-Neu) showed superoxide production, phagocytosis, bacterial killing, and chemotaxis, similar to peripheral bloo