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Differentiate hematopoietic cells from hESCs

Related

  • Stem cell resources
    • Germline stem cell markers
      • The role of microRNAs in stem cells and differentiation
          • Immunology resources
            • Immune cell markers poster
              • Cytokines for Th, Tfh, and Treg cell differentiation
                  • Protein and peptide resources
                    • Cytokines, chemokines and growth factor resources

                      The cytokines and co-culture feeder cells you need to differentiate hESCs into mature myeloid and lymphoid cells.

                      Pluripotent stem cells can be used to generate hematopoietic stem and mature cells using in vitro differentiation protocols. Here we use human embryonic stem cells (hESCs) as the source of pluripotent stem cells and highlight the relevant cytokines and feeder cells necessary to differentiate them towards myeloid and lymphoid lineages.


                      Mature cell

                      Differentiating cytokines

                      Co-culture

                      Reference

                      B cells

                      BMP4, VEGF, FGF1, bFGF, SCF, Flt3-L, TPO, GM-CSF, IL-2, IL-4, IL-15, G-CSF, IL-3, IL-6, IL-7

                      OP9

                      1

                      IL-7, IL-3, SCF, Flt3-L

                      OP9, MS5

                      2

                      Dendritic cells

                      GM-CSF, IL-4, TNF-α


                      3

                      SCF, Flt3-L, GM-CSF, IL-3, TPO, IL-4, TNF-a


                      4

                      Eosinophils

                      IL-3, IL-5

                      OP9

                      3

                      Erythrocytes

                      bFGF, VEGF, EPO, SCF, nFlt3-L, IL-3, IL-6, G-CSF, TPO


                      5

                      SCF, IL-3, IL-6, TPO, G-CSF, EPO

                      mFL stromal cells

                      6

                      EPO, TPO, IL-3, IL-6, Flt3-L, SCF, Dex

                      OP9, MS5

                      7

                      Granulocytes

                      IGF-II, VEGF, SCF, Flt3-L, TPO, G-CSF


                      8

                      BMP4, VEGF, SCF, TPO, Flt3-L, IL-3, G-CSF


                      9

                      IL-3, G-CSF or GM-CSF


                      10

                      Macrophage

                      M-CSF, IL-1β


                      3

                      Megakaryocytes/ platelets

                      BMP4, VEGF, bFGF, SCF, TPO, IL-3


                      11

                      BMP4, VEGF, IL-3, Flt3-L, TPO, SCF, EPO

                      OP9

                      12

                      BMP4, VEGF, bFGF, TPO, SCF, Flt3-L, IL-3, IL-6, IL-9


                      13

                      Neutrophil

                      SCF, Flt-3 ligand, IL-6, IL-6 receptor, thrombopoietin, IL-3, G-CSF

                      OP9

                      14

                      G-CSF

                      OP9

                      3

                      NK cells

                      BMP4, bFGF, Activin A, VEGF, IGF-1, IL-6, IL-11, SCF, IL-3, EPO, TPO, IL-13, Flt3-L, IL-15

                      OP9-DL4

                      15

                      BMP4, VEGF, SCF, IL-3, Il-6, TPO, EPO, IL-7, Flt3-L, IL-15

                      OP9-DL1

                      16

                      T cells

                      Flt-3 ligand, IL-7, SCF

                      OP9-DL1

                      17

                      BMP4, bFGF, Activin A, VEGF, IL-6, IGF-1, IL-11, SCF, EPO, TPO, Flt3-L, IL-7, IL-15

                      OP9-DL4

                      18

                      BMP4, bFGF, Activin A, VEGF, IGF-1, IL-6, IL-11, SCF, IL-3, EPO, TPO, IL-3, Flt3-L, IL-7

                      OP9-DL4

                      15

                      BMP4, bFGF, VEGF, TPO, SCF, IL-6, IL-3, IL-7, Flt3-L

                      OP9-DL1, OP9-DL4

                      19




                      After differentiation, myeloid and lymphoid cells can be characterized by flow cytometry using specific cell surface markers from our immune cell markers poster.


                      Our cytokines and growth factors


                      References

                      1.    Zambidis, E. T. et al. Expression of ACE (CD143) identifies and regulates primitive hemangioblasts derived from human pluripotent stem cells. Blood 112, 3601–3615 (2008).

                      2.    French, A., Yang, C.-T., Taylor, S., Watt, S. M. & Carpenter, L. Human Induced Pluripotent Stem Cell-Derived B Lymphocytes Express sIgM and Can be Generated via a Hemogenic Endothelium Intermediate. Stem Cells Dev. 0, 150225071446008 (2015).

                      3.    Choi, K. D., Vodyanik, M. A. & Slukvin, I. I. Generation of mature human myelomonocytic cells through expansion and differentiation of pluripotent stem cell-derived lin-CD34+CD43 +CD45+ progenitors. J. Clin. Invest. 119, 2818–2829 (2009).

                      4.    Su, Z., Frye, C., Bae, K. M., Kelley, V. & Vieweg, J. Differentiation of human embryonic stem cells into immunostimulatory dendritic cells under feeder-free culture conditions. Clin Cancer Res 14, 6207–6217 (2008).

                      5.    Chang, K. H. et al. Definitive-like erythroid cells derived from human embryonic stem cells coexpress high levels of embryonic and fetal globins with little or no adult globin. Blood 108, 1515–1523 (2006).

                      6.    Ma, F. et al. Generation of functional erythrocytes from human embryonic stem cell-derived definitive hematopoiesis. Proc. Natl. Acad. Sci. USA 105, 13087–13092 (2008).

                      7.    Dias, J. et al. Generation of red blood cells from human induced pluripotent stem cells. Stem Cells Dev. 20, 1639–47 (2011).

                      8.    Saeki, K. et al. A feeder-free and efficient production of functional neutrophils from human embryonic stem cells. Stem Cells 27, 59–67 (2009).

                      9.    Niwa, A. et al. A novel Serum-Free monolayer culture for orderly hematopoietic differentiation of human pluripotent cells via mesodermal progenitors. PLoS One 6, (2011).

                      10. Lachmann, N. et al. Large-scale hematopoietic differentiation of human induced pluripotent stem cells provides granulocytes or macrophages for cell replacement therapies. Stem Cell Reports 4, 282–296 (2015).

                      11.  Pick, M., Azzola, L., Osborne, E., Stanley, E. G. & Elefanty, A. G. Generation of Megakaryocytic Progenitors from Human Embryonic Stem Cells in a Feeder- and Serum-Free Medium. PLoS One 8, (2013).

                      12.  Vanhee, S. et al. In vitro human embryonic stem cell hematopoiesis mimics MYB-independent yolk sac hematopoiesis. Haematologica 100, 157–166 (2015).

                      13.  Feng, Q. et al. Scalable generation of universal platelets from human induced pluripotent stem cells. Stem Cell Reports 3, 817–831 (2014).

                      14.  Yokoyama, Y. et al. Derivation of functional mature neutrophils from human embryonic stem cells. Blood 113, 6584–6592 (2009).

                      15.  Sturgeon, C. M., Ditadi, A., Awong, G., Kennedy, M. & Keller, G. Wnt signaling controls the specification of definitive and primitive hematopoiesis from human pluripotent stem cells. Nat. Biotechnol. 32, 554–61 (2014).

                      16.  Ferrell, P. I., Xi, J., Ma, C., Adlakha, M. & Kaufman, D. S. The RUNX1 +24 enhancer and P1 promoter identify a unique subpopulation of hematopoietic progenitor cells derived from human pluripotent stem cells. Stem Cells 33, 1130–1141 (2015).

                      17.  Timmermans, F. et al. Generation of T cells from human embryonic stem cell-derived hematopoietic zones. J. Immunol. 182, 6879–6888 (2009).

                      18.  Kennedy, M. et al. T Lymphocyte Potential Marks the Emergence of Definitive Hematopoietic Progenitors in Human Pluripotent Stem Cell Differentiation Cultures. Cell Rep. 2, 1722–1735 (2012).

                      19.  Uenishi, G. et al. Tenascin C promotes hematoendothelial development and T lymphoid commitment from human pluripotent stem cells in chemically defined conditions. Stem Cell Reports 3, 1073–1084 (2014).

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