Methods for production of platelets from pluripotent stem cells and compositions thereof
AssigneeAstellas Institute for Regenerative Medicine
InventorsQiang Feng, Shi-Jiang Lu, Robert P. Lanza
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Abstract
Methods for production of platelets from pluripotent stem cells, such as human embryonic stem cells (hESCs) and induced pluripotent stem cells (iPSCs) are provided. These methods may be performed without forming embryoid bodies or clusters of pluripotent stem cells, and may be performed without the use of stromal inducer cells. Additionally, the yield and/or purity can be greater than has been reported for prior methods of producing platelets from pluripotent stem cells. Also provided are compositions and pharmaceutical preparations comprising platelets, preferably produced from pluripotent stem cells.
Claims32
A method for producing human megakaryocytes comprising:
- (a) culturing human pluripotent stem cells under an adherent condition in culture medium that comprises bone morphogenetic protein 4, basic fibroblast growth factor, and vascular endothelial growth factor to form human hemogenic endothelial cells;
- (b) culturing the human hemogenic endothelial cells in culture medium that comprises two or more factors selected from thrombopoietin, interleukin-3, and interleukin-6 to form human megakaryocyte progenitors;
- and
- (c) culturing the human megakaryocyte progenitors under a non-adherent culture condition in culture medium that comprises thrombopoietin and one or more factor(s) selected from stem cell factor, interleukin-6, interleukin-9, and a ROCK inhibitor to form human megakaryocytes.
A method for producing human megakaryocytes comprising:
- (a) culturing human pluripotent stem cells under an adherent condition in culture medium that comprises bone morphogenetic protein 4, basic fibroblast growth factor, and vascular endothelial growth factor to form human hemogenic endothelial cells;
- (b) culturing the human hemogenic endothelial cells in culture medium that comprises stem cell factor, thrombopoietin, Fms-related tyrosine kinase 3 ligand, interleukin-3, and interleukin-6 to form human megakaryocyte progenitors;
- and
- (c) culturing the human megakaryocyte progenitors under a non-adherent culture condition in culture medium that comprises stem cell factor, thrombopoietin, interleukin-6, and interleukin-9 to form human megakaryocytes.
A method for producing human megakaryocytes comprising:
- (a) culturing human pluripotent stem cells under an adherent condition in culture medium that comprises bone morphogenetic protein 4, basic fibroblast growth factor, and vascular endothelial growth factor to form human hemogenic endothelial cells;
- (b) culturing the human hemogenic endothelial cells in culture medium that comprises two or more factors selected from thrombopoietin, interleukin-3, interleukin-6, and Fms-related tyrosine kinase 3 to form human megakaryocyte progenitors;
- and
- (c) culturing the human megakaryocyte progenitors under a non-adherent culture condition in culture medium that comprises thrombopoietin and one or more factor(s) selected from stem cell factor, interleukin-6, interleukin-9, and a ROCK inhibitor to form human megakaryocytes.
A method for producing human megakaryocytes comprising:
- (a) culturing human pluripotent stem cells under an adherent condition in culture medium that comprises bone morphogenetic protein 4, basic fibroblast growth factor, and vascular endothelial growth factor to form human hemogenic endothelial cells;
- (b) culturing the human hemogenic endothelial cells in culture medium that comprises two or more factors selected from thrombopoietin, interleukin-3, interleukin-6, and stem cell factor to form human megakaryocyte progenitors;
- and
- (c) culturing the human megakaryocyte progenitors under a non-adherent culture condition in culture medium that comprises thrombopoietin and one or more factor(s) selected from stem cell factor, interleukin-6, interleukin-9, and a ROCK inhibitor to form human megakaryocytes.
Dependencies are inferred from the claim text.