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Stem Cells

Hematopoietic differentiation of umbilical cord blood-derived very small embryonic/epiblast-like stem cells

Leukemia volume 25, pages 1278–1285 (2011)Cite this article

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Abstract

A population of CD133+Lin−CD45− very small embryonic/epiblast-like stem cells (VSELs) has been purified by multiparameter sorting from umbilical cord blood (UCB). To speed up isolation of these cells, we employed anti-CD133-conjugated paramagnetic beads followed by staining with Aldefluor to detect aldehyde dehydrogenase (ALDH) activity; we subsequently sorted CD45−/GlyA−/CD133+/ALDHhigh and CD45−/GlyA−/CD133+/ALDHlow cells, which are enriched for VSELs, and CD45+/GlyA /CD133+/ALDHhigh and CD45+/GlyA−/CD133+/ALDHlow cells, which are enriched for hematopoietic stem/progenitor cells (HSPCs). Although freshly isolated CD45− VSELs did not grow hematopoietic colonies, the same cells, when activated/expanded over OP9 stromal support, acquired hematopoietic potential and grew colonies composed of CD45+ hematopoietic cells in methylcellulose cultures. We also observed that CD45−/GlyA−/CD133+/ALDHhigh VSELs grew colonies earlier than CD45−/GlyA−/CD133+/ALDHlow VSELs, which suggests that the latter cells need more time to acquire hematopoietic commitment. In support of this possibility, real-time polymerase chain reaction analysis confirmed that, whereas freshly isolated CD45−/GlyA−/CD133+/ALDHhigh VSELs express more hematopoietic transcripts (for example, c-myb), CD45−/GlyA−/CD133+/ALDHlow VSELs exhibit higher levels of pluripotent stem cell markers (for example, Oct-4). More importantly, hematopoietic cells derived from VSELs that were co-cultured over OP9 support were able to establish human lympho-hematopoietic chimerism in lethally irradiated non-obese diabetic/severe combined immunodeficiency mice 4–6 weeks after transplantation. Overall, our data suggest that UCB-VSELs correspond to the most primitive population of HSPCs in UCB.

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Acknowledgements

This work was supported by NIH R01 CA106281-01, NIH R01 DK074720, EU structural funds, KBN grants (N N401 024536 and N N401 0602 33), Innovative Economy Operational Program POIG.01.01.01-00-109/09-01 and the Henry M and Stella M Hoenig Endowment to MZR, the Abraham J and Phyllis Katz Foundation and the Dr Donald and Ruth Weber Goodman Philanthropic Fund (MJL). We acknowledge the Cleveland Cord Blood Center and the Cleveland Volunteer Donating Community as the source of the Material, and the efforts of the staff at the Cleveland Cord Blood Center.

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Authors and Affiliations

  1. Stem Cell Institute at the James Graham Brown Cancer Center, University of Louisville, Louisville, KY, USA

    J Ratajczak, E Zuba-Surma, I Klich, R Liu, M Wysoczynski, M Kucia & M Z Ratajczak

  2. Department of Physiology Pomeranian Medical University, Szczecin, Poland

    J Ratajczak & M Z Ratajczak

  3. Cleveland Cord Blood Center, Cleveland, OH, USA

    N Greco

  4. Hematopoietic Stem Cell Transplant Program University of Virginia, Charlottesville, VA, USA

    M J Laughlin

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  1. J Ratajczak
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Correspondence to M Z Ratajczak.

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Ratajczak, J., Zuba-Surma, E., Klich, I. et al. Hematopoietic differentiation of umbilical cord blood-derived very small embryonic/epiblast-like stem cells. Leukemia 25, 1278–1285 (2011). https://doi.org/10.1038/leu.2011.73

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