Large-Scale Hematopoietic Differentiation of Human Induced Pluripotent Stem Cells Provides Granulocytes or Macrophages for Cell Replacement Therapies

• Published in: Stem cell reports 2015-02, Vol.4 (2), p.282-296
• Main Authors: Lachmann, Nico, Ackermann, Mania, Frenzel, Eileen, Liebhaber, Steffi, Brennig, Sebastian, Happle, Christine, Hoffmann, Dirk, Klimenkova, Olga, Lüttge, Doreen, Buchegger, Theresa, Kühnel, Mark Philipp, Schambach, Axel, Janciauskiene, Sabina, Figueiredo, Constanca, Hansen, Gesine, Skokowa, Julia, Moritz, Thomas
• Format: Article
• Language: English
• Subjects: Cell Culture Techniques; Cell Differentiation - drug effects; Cell- and Tissue-Based Therapy - methods; Granulocyte Colony-Stimulating Factor - pharmacology; Granulocyte-Macrophage Colony-Stimulating Factor - pharmacology; Granulocytes - cytology; Granulocytes - metabolism; Humans; Immunohistochemistry; Immunophenotyping; Induced Pluripotent Stem Cells - cytology; Induced Pluripotent Stem Cells - drug effects; Induced Pluripotent Stem Cells - metabolism; Interleukin-3 - pharmacology; Macrophages - cytology; Macrophages - metabolism; Phenotype; Resource
• ISSN: 2213-6711; 2213-6711
• DOI: 10.1016/j.stemcr.2015.01.005

Interleukin-3 (IL-3) is capable of supporting the proliferation of a broad range of hematopoietic cell types, whereas granulocyte colony-stimulating factor (G-CSF) and macrophage CSF (M-CSF) represent critical cytokines in myeloid differentiation. When this was investigated in a pluripotent-stem-cell-based hematopoietic differentiation model, IL-3/G-CSF or IL-3/M-CSF exposure resulted in the continuous generation of myeloid cells from an intermediate myeloid-cell-forming complex containing CD34+ clonogenic progenitor cells for more than 2 months. Whereas IL-3/G-CSF directed differentiation toward CD45+CD11b+CD15+CD16+CD66b+ granulocytic cells of various differentiation stages up to a segmented morphology displaying the capacity of cytokine-directed migration, respiratory burst response, and neutrophil-extracellular-trap formation, exposure to IL-3/M-CSF resulted in CD45+CD11b+CD14+CD163+CD68+ monocyte/macrophage-type cells capable of phagocytosis and cytokine secretion. Hence, we show here that myeloid specification of human pluripotent stem cells by IL-3/G-CSF or IL-3/M-CSF allows for prolonged and large-scale production of myeloid cells, and thus is suited for cell-fate and disease-modeling studies as well as gene- and cell-therapy applications. [Display omitted] •Myeloid specification of human PSCs by IL-3-/M-CSF, G-CSF, or GM-CSF•Large-scale and continuous generation of M2-MΦ or granulocytes by M-CSF or G-CSF•Functional iPSC-derived macrophages or granulocytes similar to in-vivo-derived cells In this article, Moritz and colleagues show that IL-3 in combination with M-CSF/G-CSF or GM-CSF is able to instruct human pluripotent stem cells toward a myeloid lineage. The use of IL-3/G-CSF or IL-3/M-CSF allows for prolonged and large-scale production of human granulocytes or macrophages, respectively, that demonstrate similar functionalities compared with their in-vivo-derived counterparts.