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Reactive oxygen species levels differentiate CD34 + human progenitors based on CD38 expression
Author(s) -
Vig Christine,
Lachot Sébastien,
Foucault Amélie,
Ravalet Noémie,
Gyan Emmanuel,
Picou Frédéric,
Herault Beatrice,
Le Nail LouisRomée,
Bene Marie C.,
Herault Olivier
Publication year - 2020
Publication title -
cytometry part b: clinical cytometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.646
H-Index - 61
eISSN - 1552-4957
pISSN - 1552-4949
DOI - 10.1002/cyto.b.21948
Subject(s) - cd38 , cd34 , cord blood , progenitor cell , haematopoiesis , bone marrow , flow cytometry , stem cell , microbiology and biotechnology , biology , chemistry , immunology , andrology , medicine
Low reactive oxygen species (ROS) levels are well‐established characteristics of mouse hematopoietic stem cells (HSCs). However, little is known about these levels in human HSCs. This study aimed at quantifying ROS levels in human CD34 + CD38 low and CD34 + CD38 high human progenitors from bone marrow, cord blood and cells mobilized for autologous HSC transplantation. A specifically devised multiparameter flow cytometry method was used to quantify ROS levels (H 2 DCFDA staining) in sub‐populations of primary cells. Results were confirmed by assessing gene expression level of the ROS scavenger GPX3, a key determinant of HSC self‐renewal, in sorted CD34 + CD38 low and CD34 + CD38 high cells. CD34 + CD38 low cells from bone marrow and cord blood displayed significantly lower levels of ROS than CD34 + CD38 high cells and other leukocytes. Moreover, the correlation between ROS and GPX3 expression was verified in sorted CD34 + CD38 low and CD34 + CD38 high cells. These results confirm, in human, data previously reported in mice. Moreover, the flow cytometry assay we developed could allow for a more precise enumeration of repopulating primitive progenitors in the course of HSC transplantation.