Myeloid, B, and T lineage regeneration from pluripotent stem cells by defined transcription factors

We identified that synergistic and inducible expression of specific transcription factors in pluripotent stem cells gave rise to engraftable iHPC capable of developing into multiple lineages including myeloid cells, T cells, and B cells in the C57BL/6 recipients. The regenerative myeloid, T, and B cells were detected in multiple tissues including peripheral blood, bone marrow and spleen. We performed single cell RNA-seq on the sorted GFP+CD45.2+ cells from the peripheral blood of recipients at 16 weeks after iHPC transplantation. Single-cell transcriptome illustrated the existence of regenerative myeloid cells, T cells, and B cells. The cell hierarchy of regenerative myeloid cells, T cells, and B cells in vivo resembles those of natural counterparts. At 16 weeks after iHPC transplantation, we sorted the GFP+CD45.2+ cells from the peripheral blood of recipients for Droplet-based scRNA-seq. Droplet-based scRNA-seq datasets were produced using a Chromium system (10x Genomics) following manufacture’s instruction.