Identification and characterization of regenerative innate lymphoid cells from pluripotent stem cells [WT_ILC/R9_iILC_SmartSeq2]

We identified that synergistic and inducible expression of Runx1 and Hoxa9 in pluripotent stem cells gave rise to engraftable iHPC capable of developing into mature ILCs in the Rag2-/-Il2rg-/- recipients. We performed single-cell RNA-seq on the entire regenerative CD45.2+GFP+Lin-CD127+ R9-ILC population from the intestine of Rag2-/-Il2rg-/- recipient mice and the CD45.2+Lin-CD127+ WT-ILC population from the intestine of wild type mice (C57BL/6, CD45.2). Single-cell transcriptome illustrated the transcriptome landscape of the R9-ILCs and wild type ILCs including ILC1, ILC2 and ILC3. mRNA profiles of 67 single CD45.2+GFP+Lin-CD127+ R9-ILCs and 31 single CD45.2+Lin-CD127+ WT-ILCs. Single regenerative R9-ILCs were sorted from the intestine of Rag2-/-Il2rg-/- recipient mice 3 weeks after transplantation. Single WT-ILCs were sorted from the intestine of wild type mouse (C57BL/6, CD45.2 strain). R9-ILCs and WT-ILCs were treated according to kit instructions of Discover-sc WTA Kit V2 (Vazyme) and TruePrepTM DNA Library Prep Kit V2 (Vazyme)