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Updated: Oct 3, 2025
Identification and Isolation of Oligopotent and Lineage-committed Myeloid Progenitors from Mouse Bone Marrow
Published on: July 29, 2018
Isolation of Murine Myeloid Progenitor Populations by CD34/CD150 Surface Markers
Leonid Olender1, Roshina Thapa1, Roi Gazit1
1Shraga Segal Department of Microbiology, Immunology, and Genetics, Faculty of Health Sciences and the National Institute of Biotechnology in the Negev, Ben-Gurion University of the Negev, Beer-Sheva 84105, Israel.
Abstract:
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 FcγR 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 FcγR. We re-analyzed published single-cell RNA-Seq data and found that CD34/CD150 provides better sub-populations separation, compared to the "classical" CD34/FcγR-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.
Insights
Researchers propose a new method for separating mouse myeloid progenitor cells. Using CD34 and CD150 surface markers improves cell population distinction compared to the traditional FcγR marker.
Area of Science:
- Hematology
- Immunology
- Cell Biology
Background:
- Myeloid progenitors are crucial intermediates between hematopoietic stem cells (HSCs) and mature myeloid cells.
- In mouse bone marrow, these progenitors reside within the Lineage- cKit+ Sca1- (LK) compartment.
- Current methods using CD34 and FcγR surface markers for LK sub-population separation lack distinctness via fluorescence-activated cell sorting (FACS).
Purpose of the Study:
- To identify a more effective surface marker strategy for dissecting the mouse LK progenitor compartment.
- To evaluate CD150 as a superior alternative to FcγR for sub-dividing LK populations.
Main Methods:
- Re-analysis of published single-cell RNA-Seq data.
- Fluorescence-activated cell sorting (FACS) analysis using CD34/CD150 and CD34/FcγR gating strategies.
- Assessment of differentiation potential of newly defined LK sub-populations.
Main Results:
- Analysis of single-cell RNA-Seq data indicated that CD34/CD150 provides better sub-population separation than CD34/FcγR.
- Independent FACS analysis confirmed the improved resolution of LK sub-populations using the CD34/CD150 strategy.
- The newly identified LK sub-populations exhibited comparable differentiation potential to those defined by the classical approach.
Conclusions:
- The CD34/CD150 gating strategy offers a robust and reproducible method for separating the LK compartment into distinct myeloid progenitor sub-populations.
- This approach utilizes commonly available surface markers, facilitating broader adoption in research.
- This improved separation aids in a more precise understanding of myeloid cell development and function.
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