Lineage marker expression on mouse hematopoietic stem cells

• Published in: Experimental hematology Vol. 76; pp. 13 - 23.e2
• Main Authors: Wang, Jinhong, Liu, Zixian, Zhang, Shanshan, Wang, Xiaofang, Bai, Haitao, Xie, Miner, Dong, Fang, Ema, Hideo
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier Inc 01.08.2019
• Subjects: Animals; Antigens, CD - biosynthesis; Antigens, CD - genetics; Antigens, Differentiation - biosynthesis; Antigens, Differentiation - genetics; Bone Marrow Transplantation; Cell Lineage; Cell Self Renewal; Cell Separation; Clone Cells; Flow Cytometry; Gene Expression Regulation; Hematopoiesis; Hematopoietic Stem Cells - cytology; Hematopoietic Stem Cells - metabolism; Integrins - biosynthesis; Integrins - genetics; Mice; Mice, Inbred C57BL; Mice, Transgenic; Radiation Chimera; Thrombopoiesis
• ISSN: 0301-472X; 1873-2399
• DOI: 10.1016/j.exphem.2019.07.001

•Both Mac-1-negative and Mac-1-low HSCs exist in adult mouse bone marrow.•Both CD41-negative and CD41-low HSCs exist in adult mouse bone marrow.•No use of Mac-1 and CD41 as markers in HSC purification is recommended. Whether hematopoietic stem cells (HSCs) express lineage markers is controversial. In this study, we highly purified HSCs from the adult bone marrow of C57BL/6 mice and examined their gene expression and reconstitution potential. We first focused on the integrin family. Single-cell reverse transcription polymerase chain reaction revealed that the expression of ItgaM/Itgb2 (Mac-1) and Itga2b/Itgb3 (CD41/CD61) gradually increased along HSC differentiation, whereas Itga4, Itga5, Itga6, and ItgaV (CD51) together with Itgb1 were highly expressed in both HSCs and hematopoietic progenitor cells (HPCs). We next fractionated HSCs based on their expression of Mac-1, CD41, and CD51 by flow cytometry. We detected Mac-negative and Mac-low, but not Mac-high cells, in the HSC population. We also detected CD41-negative, -low, and -high cells in the HSC population. Competitive repopulation revealed that Mac-1-negative and -low HSCs were functionally similar, and CD41-negative and -low HSCs were functionally similar, at the single-cell level, but CD41-high HSCs were not detectable. We then found that the selection of Mac-1-negative HSCs or CD41-negative HSCs had no advantage in HSC purification. We moreover found that HSCs expressed more CD51 than CD41, and HPCs expressed more CD41 than CD51, suggesting that CD51 expression was gradually replaced by CD41 expression during megakaryocyte differentiation. We concluded that low levels of Mac-1 and CD41 expression are irrelevant to the self-renewal and differentiation potentials in HSCs.