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US 2020/0263132 A1Published application
Methods for producing enucleated erythroid cells derived from pluripotent stem cells
AssigneeAstellas Institute for Regenerative Medicine
InventorsRobert Lanza, Shi-Jiang Lu
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Abstract
Methods for generating enucleated erythroid cells using pluripotent stem cells are provided. The methods permit the production of large numbers of cells. The cells obtained by the methods disclosed may be used for a variety of research, clinical, and therapeutic applications. Methods for generating megakaryocyte and platelets are also provided.
Claims21
1-83. (canceled)
A method of producing a human pluripotent stem cell-derived megakaryocyte or a human pluripotent stem cell-derived platelet, the method comprising:
- (a) culturing a human pluripotent stem cell in a culture media comprising bone morphogenic protein 4 (BMP-4) or vascular endothelial growth factor (VEGF), or both, and inducing formation of the pluripotent stem cell into an embryoid body, in the absence of thrombopoietin (TPO), Flt-3L (FL) or stem cell factor (SCF);
- (b) culturing the embryoid body in a semi-solid culture media comprising at least two growth factors selected from the group consisting of insulin, transferrin, granulocyte macrophage colony-stimulating factor (GM-CSF), interleukin-3 (IL-3), interleukin-6 (IL-6), granulocyte colony-stimulating factor (G-CSF), stem cell factor (SCF), thrombopoietin (TPO), FLT3 (FL), vascular endothelial growth factor (VEGF), and bone morphogenic protein 4 (BMP-4), and generating a hemangioblast, a non-engrafting hemangio cell, or a blast cell from the embryoid body;
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- (c) culturing the hemangioblast, the non-engrafting hemangio cell, or the blast cell in a megakaryocyte (MK) culture media comprising TPO, thereby generating a megakaryocyte or a platelet.
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