Production of red blood cells and platelets from stem cells
This disclosure provides methods of making a megakaryocyte-erythroid progenitor cell (MEP), comprising differentiating a MEP precursor cell into a MEP in culture in the presence of an aryl hydrocarbon receptor (AhR) modulator. In some embodiments the AhR modulator is an AhR antagonist. In some embodiments the AhR modulator is an AhR agonist. In some embodiments the methods comprise culturing MEP precursor cells in the presence of an AHR antagonist and then culturing MEP precursor cells in the presence of an AHR agonist. In some embodiments the stem cell is a pluripotent stem cell. In some embodiments the MEP co-expresses CD41 and CD235. In some embodiments the number of MEPs produced in the culture increases exponentially. Methods of making a red blood cell (RBC) by culturing a MEP in the presence of an AhR modulator are also provided. Methods of making a megakaryocyte and/or a platelet, comprising culturing a MEP in the presence of an AhR modulator are also provided. In some embodiments the AhR modulator is an AhR antagonist. This disclosure also provides compositions comprising at least 1 million MEPs per ml and compositions in which at least 50% of the cells are MEPs, among other things.
1. A method of making megakaryocyte-erythroid progenitor cells (MEPs), comprising:
providing MEP precursor cells; and
culturing the MEP precursor cells in the presence of an aryl hydrocarbon receptor (AhR) antagonist for a first culture period of at least three hours and then culturing the resultant MEP precursor cells in the presence of an AHR agonist for a second culture period to make MEPs.
2. The method of claim 1 , wherein the MEP precursor cells are pluripotent stem cells.
3. The method of claim 1 , wherein the second culture period is at least 30 days; and wherein the culture continues to produce new MEP cells for the second culture period of at least 30 days.
4. The method of claim 1 , wherein the second culture period comprises 24 hours; and wherein the number of MEPs produced in the culture increases exponentially over a period of at least 24 hours during the second culture period.
5. The method of claim 1 , comprising wherein the method produces a culture comprising at least 1 million MEPs per ml.
6. The method of claim 1 , wherein the method produces a culture comprising at least 10 million MEPs per ml.
7. A method of making red blood cells (RBCs), comprising:
making MEPs according to the method of claim 1 , and
culturing the MEPs under conditions sufficient to make RBCs.
8. The method of claim 7 , wherein the conditions sufficient to make RBCs comprise culturing in erythroid specification media.
9. The method of claim 8 , wherein the conditions sufficient to make RBCs further comprise culturing in the presence of an AhR agonist.