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Impaired stem cell function of CD34+ cells selected by two different immunomagnetic beads systems

Leukemia volume 18, pages 566–574 (2004)Cite this article

Abstract

We have been investigating the hematopoietic stem cell (HSC) activity of peripheral blood-derived CD34+ cells selected by two different laboratory immunomagnetic beads systems (MiniMACS and Isolex 50). In this study, the quality of purified CD34+ cells was directly compared using clonal cell culture, a cobblestone area-forming cell (CAFC) assay, and an in vivo severe combined immunodeficiency (SCID)-repopulating cell (SRC) assay. It was found that CD34+ cells selected by these two immunomagnetic methods showed a reduced yield of colony-forming cells and CAFCs compared with cells enriched by the StemSep device (a negative selection method). However, these CD34+ cells still showed significant SRC activity, including multilineage lymphomyeloid reconstitution. The percentage of human CD45+ cells in murine bone marrow after transplanting 5 × 105 CD34+ cells selected by the Isolex 50 was significantly lower than after transplanting cells selected by the MiniMACS or the StemSep. Our findings clearly demonstrated that CD34+ cells selected by the MiniMACS system had superior HSC functions, including SRC activity, compared with cells separated by the Isolex 50 system. More detailed functional analysis of immunomagnetically separated CD34+ cells may provide useful knowledge for basic research on HSCs as well as for clinical HSC transplantation.

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Acknowledgements

This work was supported in part by Grants-in-Aid for Scientific Research on Priority areas (Grant No. 15039227) and for Scientific Research C (Grant No. 15591015) from the Ministry of Education, Science and Culture of Japan, and Japan Leukemia Research Foundation. We are grateful to Dr Hikari Nishigaki of Ohtsu Municipal Hospital for providing a sample for this study. We also thank Kirin Brewery Co., Ltd for providing the various growth factors used in this study. Furthermore, we also wish to thank Dr Ohtsura Niwa of Radiation Biology Center, Kyoto University for his advice on the irradiation of NOD/SCID mice.

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Authors and Affiliations

  1. Department of Molecular-Targeting Cancer Prevention, Division of Hematology and Oncology, Graduate School of Medical Science, Kyoto Prefectural University of Medicine, Kyoto, Japan

    T Kimura, H Minamiguchi, J Wang & Y Sonoda

  2. Second Department of Internal Medicine, Shiga University of Medical Science, Shiga, Japan

    H Minamiguchi

  3. Department of Medicine, Division of Hematology and Oncology, Graduate School of Medical Science, Kyoto Prefectural University of Medicine, Kyoto, Japan

    H Kaneko

  4. Department of Hematology, Kyoto First Red Cross Hospital, Kyoto, Japan

    H Nakagawa & H Fujii

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  1. T Kimura
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Correspondence to Y Sonoda.

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Kimura, T., Minamiguchi, H., Wang, J. et al. Impaired stem cell function of CD34+ cells selected by two different immunomagnetic beads systems. Leukemia 18, 566–574 (2004). https://doi.org/10.1038/sj.leu.2403211

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