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 stem cell into a MEP in culture in the presence of an aryl hydrocarbon receptor (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 agonist 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.
1. A method of making a platelet, comprising:
differentiating a pluripotent stem cell into a myeloid-erythroid progenitor cell (MEP) in culture in the presence of an aryl hydrocarbon receptor (AhR) agonist;
culturing the MEP under conditions sufficient to make a megakaryocyte (Mk); and
culturing the Mk under conditions sufficient for differentiation of a platelet from the Mk.
2. The method of claim 1 , wherein the conditions sufficient to make a Mk comprise culturing the MEP in the presence of an AhR antagonist.
3. The method of claim 1 , wherein the conditions sufficient to make a Mk comprise culturing in megakaryocyte specification media.
4. The method of claim 3 , wherein the conditions sufficient to make a Mk further comprise culturing in the presence of an AhR antagonist.
5. A method of making a transfusion composition, comprising making platelets by the method of claim 1 and combining the platelets with a composition comprising at least one of an anticoagulant, a buffer, and a nutrient, to thereby provide the transfusion composition.
6. A method of providing platelets to a patient in need thereof, comprising making a transfusion composition by the method of claim 5 , and transfusing the transfusion composition into the circulatory system of the patient.
7. The method of claim 6 , wherein the platelets are differentiated from induced pluripotent stem cells derived from somatic cells of the patient.
8. A method of screening a compound for an effect on platelets, comprising:
a) making platelets by the method of claim 1 ;
b) contacting the platelets with the compound; and
c) observing a change in the platelets.
9. A method of making a platelet, comprising culturing a MEP in the presence of an AhR antagonist to make a Mk and culturing the Mk under conditions sufficient for differentiation of a platelet.
10. The method of claim 9 , further comprising culturing the MEP in megakaryocyte specification media.
11. A method of making a transfusion composition, comprising making platelets by the method of claim 9 and combining the platelets with a composition comprising at least one of an anticoagulant, a buffer, and a nutrient, to thereby provide the transfusion composition.
12. A method of providing platelets to a patient in need thereof, comprising making a transfusion composition by the method of claim 11 , and transfusing the transfusion composition into the circulatory system of the patient.
13. A method of screening a compound for an effect on platelets, comprising:
a) making platelets by the method of claim 9 ;
b) contacting the platelets with the compound; and
c) observing a change in the platelets.