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Marginal expression of CXCR4 on c-kit+Sca-1+Lineage− hematopoietic stem/progenitor cells

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Abstract

Stromal cell-derived factor 1 (SDF-1) and its receptor CXCR4 are the key regulatory molecules of hematopoietic stem cell (HSC) migration and engraftment to the bone marrow (BM) microenvironment. However, the significance of the ligand–receptor complex on HSC in steady-state BM is not clear. There is currently a lack of information as to how CXCR4 is expressed on HSCs. We herein demonstrate that c-kit+Sca-1+Lineage− (KSL) cells freshly isolated from BM expressed very low to undetectable levels of CXCR4. Two hours of incubation at 37°C quickly up-modulated the receptor expression on KSL cells. Protein synthesis was not required for this early stage up-regulation, thus suggesting the emergence of intracellularly pooled receptors to the cell surface. However, protein synthesis was involved at the later stage of up-regulation. The up-regulated CXCR4 was functional, as evidenced by the fact that the incubated KSL cells more efficiently migrated to the SDF-1 gradient in vitro. Therefore, although KSL cells are able to express functional CXCR4, the receptors are only marginally expressed in the steady-state BM microenvironment. These observations therefore indicate the limited role of the SDF-1-CXCR4 axis on HSC functionality in a static BM environment.

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Acknowledgments

This work was supported by JSPS KAKENHI (No. 20591158 to Yu. Sa. No. 19591144 and No. 21591251 to Yo. So.), the Science Frontier Program from MEXT, the Twenty-first Century Center of Excellence (COE) program from MEXT, a grant from the Promotion and Mutual Aid Corporation for Private Schools of Japan, a grant from Kansai Medical University (Research grant B), a grant from the Japan Leukemia Research Foundation, a grant from the Takeda Science Foundation (to Yo. So.) and grants from Mitsubishi Pharma Research Foundation (to Yu. Sa. and Yo. So.). The authors thank Takao Kohno for his expert animal care.

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Authors and Affiliations

  1. Department of Stem Cell Biology and Regenerative Medicine, Graduate School of Medical Science, Kansai Medical University, 10-15 Fumizono-cho, Moriguchi, Osaka, 570-8506, Japan

    Yutaka Sasaki, Yoshikazu Matsuoka, Makoto Hase, Takayuki Toyohara, Mari Murakami, Masaya Takahashi, Ryusuke Nakatsuka, Yasushi Uemura & Yoshiaki Sonoda

  2. First Department of Internal Medicine, Kansai Medical University, Osaka, Japan

    Mari Murakami

  3. Department of Pediatrics, Kansai Medical University, Osaka, Japan

    Masaya Takahashi

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  1. Yutaka Sasaki
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Correspondence to Yutaka Sasaki.

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12185_2009_451_MOESM1_ESM.jpg

Supplemental Figure 1. Immunocytological detection of CXCR4. Unfractionated BM cells were visualized to express CXCR4, however, the level of CXCR4 expression on KSL cells was below the detection threshold using confocal microscopy. Of note, control antibody staining did not generate any significant background that might mask the specific staining by anti-CXCR4 antibody. The bars indicate 5 μm. (JPG 28 kb)

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Sasaki, Y., Matsuoka, Y., Hase, M. et al. Marginal expression of CXCR4 on c-kit+Sca-1+Lineage− hematopoietic stem/progenitor cells. Int J Hematol 90, 553–560 (2009). https://doi.org/10.1007/s12185-009-0451-x

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  • DOI: https://doi.org/10.1007/s12185-009-0451-x

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