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CD97 is differentially expressed on murine hematopoietic stem-and progenitor-cells

¹ Department of Immunohematology and Bloodtransfusion Leiden University Medical Center, Leiden
² Department of Experimental Immunology, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands

Correspondence: Melisssa van Pel, PhD, Section of Stem Cell Biology Department of Immunohematology and Blood Transfusion, Leiden University Medical Center, PO Box 9600, 2300 RC Leiden, The Netherlands E-mail:m.van_pel{at}lumc.nl

Background: CD97 is a member of the epidermal growth factor-seven transmembrane (EGF-TM7) family of adhesion receptors and is broadly expressed on hematopoietic cells. The aim of this study was to investigate the expression of CD97 on hematopoietic stem- and progenitor cells (HSC/HPC).

Design and Methods: CD97 expression on hematopoietic stem- and progenitor cells was studied in BALB/c, C57BL/6 and DBA/1 mice using flow cytometry. Functional hematopoietic stem- and progenitor cell characteristics were investigated in vitro and in vivo by progenitor cell assays, cobblestone area forming cell assays and bone marrow cell transplantation.

Results: Analysis of CD97 expression on murine bone marrow cells showed three major populations i.e. CD97^HI, CD97^INT and CD97^NEG cells. Functional studies revealed that radioprotective capacity and cobblestone area forming cell day 28–35 activity resides in the CD97^INT bone marrow cell fraction while CFU-GM colony-forming capacity mainly resides in the CD97^NEG population in all strains. In C57BL/6 and DBA/1 mice CD97^NEG and CD97^HI bone marrow cells show hematopoietic stem cell characteristics as well. Further functional analysis of BALB/c CD97^INT bone marrow cells revealed that c-Kit^HICD97^INT bone marrow cells exhibit HSC activity and are 1.5-fold enriched for cobblestone area forming cell-day 35 activity compared to c-Kit^HI bone marrow cells. Moreover, phenotypical analysis showed that BALB/c and C57BL/6 HSC are CD97^INT, while DBA/1 HSC are CD97^HI.

Conclusions: CD97 is differentially expressed on hematopoietic stem cells and hematopoietic progenitor cells. Committed progenitor cell activity is largely comprised in the CD97^NEG fraction, while the CD97^INT population contains hematopoietic stem cell activity. In BALB/c mice, CD97 expression can be applied to almost completely separate colony-forming cells and cells exhibiting radioprotective capacity. In addition we propose that the CD97^INTc-Kit^HI phenotype allows simple and rapid purification of murine hematopoietic stem cells.

Key words: hematopoietic stem cells, CD97, mouse model, hematopoietic progenitor cells.