Journal
CELLS
Volume 11, Issue 3, Pages -Publisher
MDPI
DOI: 10.3390/cells11030350
Keywords
myeloid progenitors; murine bone marrow; cell sorting; hematopoiesis
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In this study, the researchers propose using CD34/CD150 instead of the traditional CD34/Fc gamma R approach to separate sub-populations in the LK compartment. They re-analyzed single-cell RNA-Seq data and confirmed their findings through independent FACS analysis. The newly-obtained LK sub-populations showed comparable differentiation potential to the classical ones.
Myeloid progenitors are intermediates between Hematopoietic Stem Cells (HSCs) and Myeloid effector progeny. In mouse bone marrow, they are part of the Lineage(-) cKit(+) Sca1(-) (LK) compartment. To date, most researchers used CD34 and Fc gamma R surface markers for the dissection of this compartment into various populations. Surprisingly, however, this approach does not provide distinct separation by fluorescence-activated cell sorting (FACS). In this study, we suggest using CD150 instead of Fc gamma R. We re-analyzed published single-cell RNA-Seq data and found that CD34/CD150 provides better sub-populations separation, compared to the classical CD34/Fc gamma R-based approach. We confirm our findings by independent FACS analysis. We demonstrate comparable differentiation potential of the newly-obtained LK sub-populations, like previous classical ones. Therefore, we suggest the CD34/CD150 gating strategy, utilizing commonly-used surface markers, as a robust and reproducible separation of the LK compartment into distinct sub-populations.
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