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Human CD34-negative Hematopoietic Stem Cells

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Adult Stem Cell Therapies: Alternatives to Plasticity

Part of the book series: Stem Cell Biology and Regenerative Medicine ((STEMCELL))

Abstract

Based on the recent development of fluorescence-activated cell sorting (FACS) technology, murine hematopoietic stem cells (HSCs) can be purified at the single cell level. The immunophenotype of murine HSCs is CD34low/−c-kit+Sca-1+Lin− (CD34− KSL) cells. However, the characterization of primitive human HSCs has not been fully elucidated. The biology of human HSCs is a current topic of interest that has important implications for clinical HSC transplantation as well as basic research on HSCs. Recently, human cord blood (CB)-derived CD34− HSCs, a counterpart of murine CD34low/− KSL cells, were successfully identified using an intra-bone marrow injection (IBMI) method. This review aims to update the concept of the immunophenotype and functional characteristics of human primitive CD34− HSCs. In addition, the significance of the application of the IBMI technique in clinical CB stem cell transplantation is also discussed. Recent rapid advances in understanding the biological nature of HSCs may make it possible to fully characterize the most primitive class of human HSCs, thereby clarifying the human HSC hierarchy, in the near future.

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Authors and Affiliations

  1. Department of Stem Cell Biology and Regenerative Medicine, Graduate School of Medical Science, Kansai Medical University, Shinmachi, Hirakata, 573-1010, Osaka, Japan

    Yoshiaki Sonoda MD, PhD

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Editors and Affiliations

  1. University of Louisville, Stem Cell Institute, James Graham Brown Cancer Center, Louisville, Kentucky, USA

    Mariusz Z. Ratajczak

Conclusions and Future Prospects

Conclusions and Future Prospects

The relationship of primitive HSCs within the human HSC hierarchy has been difficult to clarify due to the heterogeneity of the stem cell compartment. This heterogeneity results in major problems in the isolation/purification of most primitive human HSCs. We successfully developed a sensitive SRC assay system using the IBMI technique to identify a new class of CB-derived CD34− HSCs [13, 27, 31, 38, 51]. Based on our current data obtained using flow cytometry and serial transplantation analyses, we propose that the surface immunophenotype of the most primitive human LTR-HSCs is Lin−CD34−FLT3−c-kit−/lowTie2−KDR−Thy1low/−CD49flow/−CD45RA−CD110−CD133+ (Fig. 4.8). However, the frequency of CB-derived CD34− SRCs is 1/142 in 18Lin−CD34−CD133+ cells [51] at the present time, which is still low in comparison to that of CD34+CD38− SRCs (1/40 cells) (31). In order to more effectively enrich/purify human CD34− LTR-HSCs at the single cell level, it is very important to identify another reliable positive marker for human LTR-HSCs beside CD133. Our goal is to achieve the complete purification of most primitive human HSCs, enabling the development of stem cell-based cellular and genetic therapies for various hematological and non-hematological diseases.

The molecular mechanisms that regulate the self-renewal and/or differentiation of human HSPCs are poorly understood because our understanding of these mechanisms, which control hematopoiesis, has primarily been obtained from mouse models. However, the cellular properties of murine and human HSCs appear to differ. Therefore, these analyses should be performed using purified human HSCs to confirm the data obtained from mouse models.

The application of our sensitive SRC assay system using the IBMI technique may make it possible to discover other hitherto unidentified HSCs in various organs and/or identify new markers for LTR-HSCs. It is also important to clarify whether the BM or mobilized PB contains an equivalent class of CD34− SRCs. From another point of view, the candidate positive markers for primitive human HSCs, such as CD133 and MPL, are expressed differently on the surface of CB- and BM-derived 18Lin−CD34− cells [51](Matsuoka and Sonoda, unpublished data). These results suggest that human CB and BM contain different classes of HSCs. As described in this review, CD34− HSCs continue to be heterogeneous populations and are not completely purified at the single cell level. Therefore, an important issue, the developmental origin of these primitive CD34−HSCs, has not been fully clarified and requires further investigation.

Acknowledgements

The author is grateful to Drs. Y. Matsuoka, M. Takahashi, R. Nakatsuka, K. Sumide, T. Fujioka, H. Kohno, and Y. Sasaki, for their contributions. The author also thanks Kirin Brewery Co. Ltd. (Tokyo, Japan) for providing the various growth factors. Ms. K. Ishino is also acknowledged for her valuable assistance in preparing the manuscript.

This work was supported by Grants-in-Aid for Scientific Research on Priority Areas (Grant No. 15039227) and for Scientific Research C (Grant Nos. 15591015, 19591144, 21591251, and 24591432) from the Ministry of Education, Science and Culture of Japan, a grant from Haiteku Research Center of the Ministry of Education, a grant from the Science Frontier Program of the Ministry of Education, a grant from the Twenty-first Century Center of Excellence (COE) program of the Ministry of Education, a grant from the Promotion and Mutual Aid Corporation for Private Schools of Japan, a grant from Kansai Medical University (Research grant B), grants from MEXT-supported Program for the Strategic Research Foundation at Private Universities (2011–2016 and 2012–2017), a grant from the Adaptable and Seamless Technology Transfer Program through target-driven R&D, JST, a grant fromthe Japan Leukemia Research Foundation, a grant from the Mitsubishi Pharma Research Foundation, a grant from the Terumo Life Science Foundation, and a grant from the SENSHIN Medical Research Foundation.

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Sonoda, Y. (2014). Human CD34-negative Hematopoietic Stem Cells. In: Ratajczak, M. (eds) Adult Stem Cell Therapies: Alternatives to Plasticity. Stem Cell Biology and Regenerative Medicine. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1001-4_4

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