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Insights into the remarkable engraftment potential of fetal liver (FL) hematopoietic stem cells (HSCs) could be harnessed to improve ex vivo HSC expansion, increase engraftment efficiency, and enable in vitro HSC generation. Using CITE-seq, a technique that combines single cell RNA sequencing with oligo-tagged antibodies, we profiled 26,407 FL cells at both the transcriptional and protein level. In these studies, we have analyzed CD34- cells, CD34+ cells and >7,000 CD34+ cells further enriched by expression of GPI-80, a marker tightly linked to engraftment potential, to explicitly characterize HSCs capable of long-term engraftment.

Superior engraftment potential of the GPI-80+ fraction confirmed enrichment for bona fide HSCs and coincided with enrichment for known HSC markers such as ITGA6 (CD49f), PROCR (CD201), MLLT3, HLF and HMGA2. Further dissection of this engraftment profile revealed …