Patent ID: 8846395
Filing Date: 2014-09-30
Classification: C12N

Abstract:
1. 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 (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.