Differentiate hematopoietic cells from hESCs
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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.
References
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