Generation of specific hematopoietic lineages from murine ES cells
Hematopoietic lineage . | Method of culture . | Cytokines . | References . | Other . |
|---|---|---|---|---|
| Megakaryocytes and platelets | OP9, EBs | Major: Tpo | 38, 115-119 | Megakaryocytes show agonist-dependent fibrinogen binding. Platelets exhibit agonist-dependent fibrinogen binding, aggregation, and spreading. |
| Other: IL-6, IL-11 | ||||
| Erythroid | OP9, EBs | Major: Epo, KL | 38, 45, 47, 114, 120 | EryPs are KL independent. EryDs require Epo/KL. Iron-saturated transferrin added to improve hemoglobin synthesis during terminal maturation. Dexamethasone can be used for erythroid progenitor expansion. |
| Other: Insulin, IGF-1, IL-3 | ||||
| Mast cells | EBs | IL-3, KL | 121, 122 | EB-derived mast cells proliferate extensively in culture and reconstitute mast cell-deficient KitW/KitW-v mice after adoptive transfer. |
| Macrophages | OP9, EBs | IL-1, IL-3, M-CSF, GM-CSF | 47, 123-125 | Macrophage overgrowth tends to inhibit the production of other lineages in ES-cell differentiation cultures. |
| T lymphocytes | OP9, OP9-DL1 | Flt-3 ligand, IL-7 | 126, 127 | Full T-lymphocyte development requires maturation in thymic stroma (FTOCs or RTOCs). T-lymphocyte production is augmented by cultivation on OP9 cells ectopically expressing the notch ligand DL-1. |
| B lymphocytes | OP9 | Flt-3 ligand, IL-7 | 47, 128, 129 | Stimulation with LPS augments production of IgM+-secreting B lymphocytes. |
| Eosinophils | OP9 | IL-5, IL-3, GM-CSF, eotaxin | 130 | Express characteristic granules and cell-surface markers, produce oxidative response to Leishmania. |
| Neutrophils | EBs, OP9 | Progenitor expansion: OSM, b-FGF, IL-6, IL-11, LIF, KL | 131, 132 | Functional neutrophils produced. This includes characteristic markers and morphology, superoxide production, and chemotactic responses. |
| Neutrophil differentiation: G-CSF, GM-CSF, IL-6 | ||||
| DCs | EBs, OP9 | Progenitor expansion: GM-CSF, IL-3 | 133-135 | DC production facilitated by plating EBs on tissue culture dishes. Exhibit characteristic surface phenotype, process protein antigens, and stimulate T lymphocytes in the MLR. |
| Differentiation/maturation: IL-4, TNF-α, LPS, anti-CD40 | ||||
| NK cells | OP9 | Flt-3 ligand, IL-15, IL-6, IL-7, KL | 129, 136 | Express characteristic NK proteins; kill selected tumor lines and MHC 1-deficient lymphoblasts. |
| Osteoclasts | OP9, ST2 | M-CSF, RANKL | 137-141 | Recognized by TRAP expression and bone resorbtion activity. Production augmented by vitamins C and D and by dexamethasone. |
Hematopoietic lineage . | Method of culture . | Cytokines . | References . | Other . |
|---|---|---|---|---|
| Megakaryocytes and platelets | OP9, EBs | Major: Tpo | 38, 115-119 | Megakaryocytes show agonist-dependent fibrinogen binding. Platelets exhibit agonist-dependent fibrinogen binding, aggregation, and spreading. |
| Other: IL-6, IL-11 | ||||
| Erythroid | OP9, EBs | Major: Epo, KL | 38, 45, 47, 114, 120 | EryPs are KL independent. EryDs require Epo/KL. Iron-saturated transferrin added to improve hemoglobin synthesis during terminal maturation. Dexamethasone can be used for erythroid progenitor expansion. |
| Other: Insulin, IGF-1, IL-3 | ||||
| Mast cells | EBs | IL-3, KL | 121, 122 | EB-derived mast cells proliferate extensively in culture and reconstitute mast cell-deficient KitW/KitW-v mice after adoptive transfer. |
| Macrophages | OP9, EBs | IL-1, IL-3, M-CSF, GM-CSF | 47, 123-125 | Macrophage overgrowth tends to inhibit the production of other lineages in ES-cell differentiation cultures. |
| T lymphocytes | OP9, OP9-DL1 | Flt-3 ligand, IL-7 | 126, 127 | Full T-lymphocyte development requires maturation in thymic stroma (FTOCs or RTOCs). T-lymphocyte production is augmented by cultivation on OP9 cells ectopically expressing the notch ligand DL-1. |
| B lymphocytes | OP9 | Flt-3 ligand, IL-7 | 47, 128, 129 | Stimulation with LPS augments production of IgM+-secreting B lymphocytes. |
| Eosinophils | OP9 | IL-5, IL-3, GM-CSF, eotaxin | 130 | Express characteristic granules and cell-surface markers, produce oxidative response to Leishmania. |
| Neutrophils | EBs, OP9 | Progenitor expansion: OSM, b-FGF, IL-6, IL-11, LIF, KL | 131, 132 | Functional neutrophils produced. This includes characteristic markers and morphology, superoxide production, and chemotactic responses. |
| Neutrophil differentiation: G-CSF, GM-CSF, IL-6 | ||||
| DCs | EBs, OP9 | Progenitor expansion: GM-CSF, IL-3 | 133-135 | DC production facilitated by plating EBs on tissue culture dishes. Exhibit characteristic surface phenotype, process protein antigens, and stimulate T lymphocytes in the MLR. |
| Differentiation/maturation: IL-4, TNF-α, LPS, anti-CD40 | ||||
| NK cells | OP9 | Flt-3 ligand, IL-15, IL-6, IL-7, KL | 129, 136 | Express characteristic NK proteins; kill selected tumor lines and MHC 1-deficient lymphoblasts. |
| Osteoclasts | OP9, ST2 | M-CSF, RANKL | 137-141 | Recognized by TRAP expression and bone resorbtion activity. Production augmented by vitamins C and D and by dexamethasone. |
DCs indicates dendritic cells; NK, natural killer; OP9, M-CSF-/- stromal line; EB, embryoid bodies; Tpo, thrombopoietin; IL-6, interleukin 6; Epo, erythropoietin; KL, c-kit ligand; IGF-1, insulin-like growth factor 1; EryP, primitive erythroid progenitor; EryD, definitive erythroid progenitor; M-CSF, macrophage colony-stimulating factor; GM-CSF granulocyte/macrophage colony-stimulating factor; FTOCs, fetal thymic organ cultures; RTOCs, reaggregate thymic organ cultures; DL-1, Delta-like-1; LPS, lipopolysaccharide; OSM, oncostatin M; b-FGF, basic fibroblast growth factor; LIF, leukemia inhibitory factor; G-CSF, granulocyte colony-stimulating factor; TNF-α, tumor necrosis factor α; MLR, mixed lymphocyte reaction; ST2, bone marrow-derived stromal line; and TRAP, tartrate-resistant acid phosphatase.