Production of Induced Pluripotent Stem Cell-Derived Red Blood Cells

Red blood cell (RBC) transfusion is one of the most common medical treatments, with over 10 million units transfused per year in the United States alone. One complication following transfusion is red cell alloimmunization, the production of antibodies to foreign proteins on donor RBCs. Identifying compatible RBC units for alloimmunized patients may be challenging due to the lack of specific RBC reagents with uncommon or rare RBC antigen phenotypes that are needed to identify the precise antibody target. Human induced pluripotent stem cells (iPSCs) represent a renewable source of cells to generate specialized RBCs by reprogramming rare donor cells or by gene editing iPSCs. These iPSCs can provide an unlimited supply of RBCs with unique antigen expression profiles. Here, we outline protocols to generate developmentally primitive (embryonic) and definitive (fetal) RBCs from iPSCs, and methods to perform antibody identification using iPSC-derived RBCs as reagent cells in gel agglutination assays commonly used by blood banks and reference immunohematology laboratories.