Mapping Human Pluripotent Stem Cell Derived Erythroid Differentiation by Single-Cell Transcriptome Analysis

Abstract

There is currently an imbalance between the supply and demand of functional red blood cells (RBCs) in clinical applications, and this imbalance can be addressed by regenerating RBCs with a variety of in vitro methods. Induced pluripotent stem cells (iPSCs) can address the low supply of cord blood and the ethical issues in embryonic stem cell research and provide a promising strategy to eliminate immune rejection. However, no complete single-cell level differentiation pathway exists for the iPSC-derived RBC differentiation system. In this study, we used iPSC line BC1 to establish an RBC regeneration system and used the 10× Genomics single-cell transcriptome platform to map the cell lineage and differentiation trajectories on day 14 (D14) of the regeneration system. We found iPSC differentiation was not synchronized during embryoid body (EB) culture, and the D14 cells in the system mainly consisted of mesodermal and various blood cells, similar to yolk sac hematopoiesis. During asynchronous EB differentiation, iPSCs undergo three bifurcations before they enter erythroid differentiation, and the driver genes of each bifurcation were identified. The key roles of cell adhesion and estradiol in RBC regeneration were observed. This study provides systematically theoretical guidance for the optimization of the iPSC-derived RBC differentiation system.