Method for producing dendritic cells from pluripotent stem cells
First Claim
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1. A method for producing dendritic cells from mammalian pluripotent stem cells, said method comprising culturing pluripotent stem cells by the steps below:
- (1) culturing the pluripotent stem cells in an adherent culture comprising a medium which comprises bone morphogenic protein 4 (BMP4) but does not comprise serum;
(2) culturing the cells obtained by step (1) in an adherent culture comprising a medium which comprises vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF) and/or stem cell factor (SCF) but does not comprise serum;
(3) culturing the cells obtained by step (2) in an adherent culture comprising a medium which comprises a hematopoietic factor but does not comprise serum, wherein said culturing in step (3) is performed in a first medium comprising hematopoietic factors consisting of (i) SCF, thrombopoietin (TPO), interleukin-3 (IL-3) and Fms-like tyrosine kinase 3 ligand (Flt3-ligand), and then in a second medium comprising hematopoietic factors consisting of (ii) granulocyte macrophage-colony stimulating factor (GM-CSF), macrophage-colony stimulating factor (M-CSF) and Flt3-ligand; and
(4) culturing the cells obtained by step (3) in a suspension culture comprising a medium which comprises GM-CSF and interleukin-4 (IL-4) but does not comprise serum, wherein dendritic cells are produced/collected.
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Abstract
A dendritic cell is produced from pluripotent stem cells by culturing pluripotent stem cells by the following steps: (1) performing adherent culture in a medium which comprises a BMP family protein but does not comprise serum; (2) performing adherent culture in a medium which comprises VEGF but does not comprise serum; (3) performing adherent culture in a medium which comprises a hematopoietic factor but does not comprise serum; and (4) performing suspension culture in a medium which does not comprise serum.
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Citations
18 Claims
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1. A method for producing dendritic cells from mammalian pluripotent stem cells, said method comprising culturing pluripotent stem cells by the steps below:
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(1) culturing the pluripotent stem cells in an adherent culture comprising a medium which comprises bone morphogenic protein 4 (BMP4) but does not comprise serum; (2) culturing the cells obtained by step (1) in an adherent culture comprising a medium which comprises vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF) and/or stem cell factor (SCF) but does not comprise serum; (3) culturing the cells obtained by step (2) in an adherent culture comprising a medium which comprises a hematopoietic factor but does not comprise serum, wherein said culturing in step (3) is performed in a first medium comprising hematopoietic factors consisting of (i) SCF, thrombopoietin (TPO), interleukin-3 (IL-3) and Fms-like tyrosine kinase 3 ligand (Flt3-ligand), and then in a second medium comprising hematopoietic factors consisting of (ii) granulocyte macrophage-colony stimulating factor (GM-CSF), macrophage-colony stimulating factor (M-CSF) and Flt3-ligand; and (4) culturing the cells obtained by step (3) in a suspension culture comprising a medium which comprises GM-CSF and interleukin-4 (IL-4) but does not comprise serum, wherein dendritic cells are produced/collected. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9)
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10. A method for producing dendritic cells from mammalian pluripotent stem cells, said method comprising culturing pluripotent stem cells by the steps below:
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(1) culturing the pluripotent stem cells in an adherent culture comprising a medium which comprises bone morphogenic protein 4 (BMP4) but does not comprise serum; (2) culturing the cells obtained by step (1) in an adherent culture comprising a medium which comprises vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF) and/or stem cell factor (SCF) but does not comprise serum; (3) culturing the cells obtained by step (2) in an adherent culture comprising a medium which comprises a hematopoietic factor but does not comprise serum, wherein said culturing in step (3) is performed in a first medium comprising hematopoietic factors consisting of (i) SCF, thrombopoietin (TPO), interleukin-3 (IL-3) and Fms-like tyrosine kinase 3 ligand (Flt3-ligand), and then in a second medium comprising hematopoietic factors consisting of (ii) granulocyte macrophage-colony stimulating factor (GM-CSF), macrophage-colony stimulating factor (M-CSF), Flt3-ligand, and SCF; and (4) culturing the cells obtained by step (3) in a suspension culture comprising a medium which comprises GM-CSF and interleukin-4 (IL-4) but does not comprise serum, wherein dendritic cells are produced/collected. - View Dependent Claims (11, 12, 13, 14, 15, 16, 17, 18)
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