Generation of mature myelomonocytic cells through expansion and differentiation of pluripotent stem cell-derived lin-CD34+CD43+CD45+progenitors

A method of producing myeloid lineage cells, comprising the steps of (a) obtaining a population of mammalian pluripotent stem cells, wherein the stem cells are derived from mammalian embryos or are induced pluripotent stem cells, (b) inducing hematopoietic differentiation to obtain lin−CD34+CD43+CD45+CD38− multipotent hematopoietic cells by co-culturing the mammalian pluripotent stem cells with stromal cells, (c) expanding and differentiating lin−CD34+CD43+CD45+CD38− cells from step (b) by culture in the presence of GM-CSF to obtain CD34+CD43+CD45+CD38+ myeloid progenitors and

• (d) exposing the CD34+CD43+CD45+CD38+ myeloid progenitors resulting from step (c) to:
• (i) G-CSF for 7-9 days, wherein a population of neutrophils is isolated;
• (ii) IL-3 and IL-5 for 12-14 days wherein a population of eosinophils is isolated;
• (iii) M-CSF and IL-1β for 1-8 days wherein a population of macrophages is isolated;
• (iv) GM-CSF, lα,25-dihydroxyvitamin D3 for 5-7 days, then GM-CSF, lα,25-dihydroxyvitamin D3 and RANKL for 7-14 days, wherein populations of osteoclasts are isolated;
• (v) GM-CSF and IL-4 for 7-9 days, TNF-α being added after day 3 of GM-CSF and IL-4 exposure, wherein a population of dendritic cells is isolated;
• or (vi) GM-CSF and TGF-β, TNF-α being added after day 3 of GM-CSF and TGF-β exposure, wherein a population of Langerhans cells is isolated.