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Isolation of Murine Myeloid Progenitor Populations by CD34/CD150 Surface Markers

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...

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Autores principales: Olender, Leonid, Thapa, Roshina, Gazit, Roi
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8834359/
https://www.ncbi.nlm.nih.gov/pubmed/35159159
http://dx.doi.org/10.3390/cells11030350
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author Olender, Leonid
Thapa, Roshina
Gazit, Roi
author_facet Olender, Leonid
Thapa, Roshina
Gazit, Roi
author_sort Olender, Leonid
collection PubMed
description 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.
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spelling pubmed-88343592022-02-12 Isolation of Murine Myeloid Progenitor Populations by CD34/CD150 Surface Markers Olender, Leonid Thapa, Roshina Gazit, Roi Cells Communication 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. MDPI 2022-01-20 /pmc/articles/PMC8834359/ /pubmed/35159159 http://dx.doi.org/10.3390/cells11030350 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Communication
Olender, Leonid
Thapa, Roshina
Gazit, Roi
Isolation of Murine Myeloid Progenitor Populations by CD34/CD150 Surface Markers
title Isolation of Murine Myeloid Progenitor Populations by CD34/CD150 Surface Markers
title_full Isolation of Murine Myeloid Progenitor Populations by CD34/CD150 Surface Markers
title_fullStr Isolation of Murine Myeloid Progenitor Populations by CD34/CD150 Surface Markers
title_full_unstemmed Isolation of Murine Myeloid Progenitor Populations by CD34/CD150 Surface Markers
title_short Isolation of Murine Myeloid Progenitor Populations by CD34/CD150 Surface Markers
title_sort isolation of murine myeloid progenitor populations by cd34/cd150 surface markers
topic Communication
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8834359/
https://www.ncbi.nlm.nih.gov/pubmed/35159159
http://dx.doi.org/10.3390/cells11030350
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