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Stem cell biology

Activation of OCT4 enhances ex vivo expansion of human cord blood hematopoietic stem and progenitor cells by regulating HOXB4 expression

Leukemia volume 30, pages 144–153 (2016)Cite this article

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Abstract

Although hematopoietic stem cells (HSC) are the best characterized and the most clinically used adult stem cells, efforts are still needed to understand how to best ex vivo expand these cells. Here we present our unexpected finding that OCT4 is involved in the enhancement of cytokine-induced expansion capabilities of human cord blood (CB) HSC. Activation of OCT4 by Oct4-activating compound 1 (OAC1) in CB CD34+ cells enhanced ex vivo expansion of HSC, as determined by a rigorously defined set of markers for human HSC, and in vivo short-term and long-term repopulating ability in NSG mice. Limiting dilution analysis revealed that OAC1 treatment resulted in 3.5-fold increase in the number of SCID repopulating cells (SRCs) compared with that in day 0 uncultured CD34+ cells and 6.3-fold increase compared with that in cells treated with control vehicle. Hematopoietic progenitor cells, as assessed by in vitro colony formation, were also enhanced. Furthermore, we showed that OAC1 treatment led to OCT4-mediated upregulation of HOXB4. Consistently, siRNA-mediated knockdown of HOXB4 expression suppressed effects of OAC1 on ex vivo expansion of HSC. Our study has identified the OCT4-HOXB4 axis in ex vivo expansion of human CB HSC.

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Acknowledgements

We sincerely thank other lab members in Dr Broxmeyer's laboratory for helpful discussion. This work was supported by Public Health Service Grants from the NIH to HEB: R01 HL056416, R01 HL67384, R01 HL112669 and NIH P01 DK090948. The teratoma study was supported by grant HI14C3365 from the Ministry for Health & Welfare by the Korean government.

Author contributions

XH and HEB directed the research, designed and performed experiments, interpreted data and wrote the manuscript. SC and GH performed experiments and interpreted data. ML, KH and HC performed the teratoma study and interpreted data.

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Authors and Affiliations

  1. Department of Microbiology and Immunology, Indiana University School of Medicine, Indianapolis, IN, USA

    X Huang, M-R Lee, S Cooper, G Hangoc & H E Broxmeyer

  2. Soonchunhyang Institute of Medi-bio Science, Chungcheongnam-do, Korea

    M-R Lee

  3. Konkuk University School of Medicine, Seoul, Korea

    K-S Hong & H-M Chung

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  1. X Huang
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  2. M-R Lee
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  4. G Hangoc
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  7. H E Broxmeyer
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Correspondence to H E Broxmeyer.

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Competing interests

HEB is a member of the Medical Scientific Advisory Board of CordUse, a cord blood banking company based in Orlando, Florida.

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Huang, X., Lee, MR., Cooper, S. et al. Activation of OCT4 enhances ex vivo expansion of human cord blood hematopoietic stem and progenitor cells by regulating HOXB4 expression. Leukemia 30, 144–153 (2016). https://doi.org/10.1038/leu.2015.189

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