Regeneration of immunocompetent B lymphopoiesis from pluripotent stem cells guided by transcription factors

We identified that the coordinated expression of exogenous transcription factors(Lhx2, Hoxa9, and Runx1), efficiently guides PSC-derived hematopoietic progenitors (iHPC) preferentially towards B cell lineage commitment, which successfully regenerate B lymphopoiesis in B-cell deficient animals. Of note, the abundancy of in vivo lymphopoiesis includes pro/pre-B progenitors, immature B cells, and all subsets of mature B1a, B1b, FO B and MZ B reconstitution in vivo. We performed single-cell RNA-Seq using the sorted GFP+CD45+ Lin- cells from the bone marrow of recipients at day 7.5 after iHPC transplantation. Overall design: At day 7.5 after iHPC transplantation, we sorted the GFP+CD45+ Lin- cells from the bone marrow of recipients for Droplet-based scRNA-seq. Droplet-based scRNA-seq datasets were produced using a Chromium system (10x Genomics, PN120263) following manufacture's instruction.