Identification and characterization of regenerative innate lymphoid cells from pluripotent stem cells [WT-CHILP/iR9-CHILP_SmartSeq2]

We identified that synergistic and inducible expression of Runx1 and Hoxa9 in pluripotent stem cells (PSCs) gave rise to engraftable iHPC capable of developing into common helper innate lymphoid progenitors (CHILPs) in the Rag2-/-Il2rg-/- recipients. To assess the gene-expression pattern of the PSC-derived R9-iCHILPs, we performed single-cell RNA-seq on the regenerative CD45.2+GFP+Lin-CD127+CD135-a4ß7+Sca1mid/-CD25- R9-iCHILPs from the bone marrow of Rag2-/-Il2rg-/- recipient mice and the CD45.2+Lin-CD127+CD135-a4ß7+Sca1mid/-CD25- WT-CHILPs from the bone marrow of wild type mice (C57BL/6, CD45.2). Overall design: mRNA profiles of 34 single CD45.2+GFP+Lin-CD127+CD135-a4ß7+Sca1mid/-CD25- R9-iCHILPs and 29 single CD45.2+Lin-CD127+CD135-a4ß7+Sca1mid/-CD25- WT-CHILPs. Single R9-iCHILP were sorted from the bone marrow of Rag2-/-Il2rg-/- recipient mice four weeks after transplantation and were treated according to kit instructions of Discover-sc WTA Kit V2 (Vazyme) and TruePrepTM DNA Library Prep Kit V2 (Vazyme).