Functional heterogeneity of mouse fetal liver hematopoietic stem cells revealed by clonal lineage tracing and single-cell transcriptomics

After confirming long‑term multilineage reconstitution, we sorted GFP⁺ hematopoietic stem and progenitor cells from the bone marrow of primary recipients. Half of the sorted cells were processed for single‑cell RNA sequencing, while the remaining HSCs were injected into secondary recipients to assess self‑renewal potential.

Following long‑term engraftment in secondary recipients, GFP⁺ HSPCs were similarly isolated. For both primary and secondary transplantation cohorts, a defined mixture of LT‑HSCs and other progenitor populations (MPPs, MyPs, MkPs, CLPs) was prepared to reconstitute the full hematopoietic stem and progenitor compartment. In total, samples from five primary and five secondary recipients were used to generate the dataset.