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Hematopoietic progenitor cell lines with myeloid and lymphoid potential

Nature Methods volume 10, pages 795–803 (2013)Cite this article

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Abstract

Investigation of immune-cell differentiation and function is limited by shortcomings of suitable and scalable experimental systems. Here we show that retroviral delivery of an estrogen-regulated form of Hoxb8 into mouse bone marrow cells can be used along with Flt3 ligand to conditionally immortalize early hematopoietic progenitor cells (Hoxb8-FL cells). Hoxb8-FL cells have lost self-renewal capacity and potential to differentiate into megakaryocytes and erythrocytes but retain the potential to differentiate into myeloid and lymphoid cells. They differentiate in vitro and in vivo into macrophages, granulocytes, dendritic cells, B lymphocytes and T lymphocytes that are phenotypically and functionally indistinguishable from their primary counterparts. Quantitative in vitro assays indicate that myeloid and B-cell potential of Hoxb8-FL cells is comparable to that of primary lymphoid-primed multipotent progenitors, whereas T-cell potential is diminished. The simplicity of this system and the unlimited proliferative capacity of Hoxb8-FL cells will enable studies of immune-cell differentiation and function.

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Figure 1: Growth and morphology of Hoxb8-FL cells.
Figure 2: Phenotype and function of Hoxb8-FL–derived myeloid cells.
Figure 3: In vivo differentiation of Hoxb8-FL cells.
Figure 4: In vivo differentiation and function of Hoxb8-FL–derived lymphocytes.
Figure 5: Hoxb8-FL cells sustain multilineage potential in vitro, but lack the potential to differentiate into megakaryocytes and erythroid cells (MkE potential).
Figure 6: Gene expression and biochemical application of Hoxb8-FL cells.

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Acknowledgements

We thank the staff of the Animal Resource Center, the Flow Cytometry and Cell Sorting Resource, and the Proteomics and Mass Spectrometry laboratory at the St. Jude Children's Research Hospital and L. Chi for technical assistance; and H. Hochrein for helpful discussion related to dendritic cell biology. Supported by National Institutes of Health National Institute of Allergy and Infectious Diseases grant AI083443 to H.H., the National Institutes of Health National Cancer Institute grant P30CA021765 and the American Lebanese Syrian Associated Charities. We thank M. Kamps (Univ. California, San Diego) for the SCF-producing B16 melanoma cell line, R. Steinman (Rockefeller Univ.) for the Flt3L-producing B16 melanoma cell line, S. Zandi and M. Sigvardsson (Linköping Univ.) and D. Bryder (Lund Univ.) for hematopoietic progenitor cell markers, and W. Goebel and D. Loeffler (Univ. Würzburg) for ovalbumin-encoding cDNA.

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Authors and Affiliations

  1. Department of Infectious Diseases, St. Jude Children's Research Hospital, Memphis, Tennessee, USA.,

    Vanessa Redecke, Ruiqiong Wu, Jingran Zhou, Vandana Chaturvedi & Hans Häcker

  2. Department of Computational Biology, St. Jude Children's Research Hospital, Memphis, Tennessee, USA

    David Finkelstein

  3. Proteomics Core Facility, St. Jude Children's Research Hospital, Tennessee, USA

    Anthony A High

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  1. Vanessa Redecke
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Contributions

V.R., R.W., V.C. and H.H. planned experiments; V.R., R.W., J.Z., V.C. and H.H. performed experiments; A.A.H., D.F., V.C., V.R. and H.H. analyzed data; V.R. and H.H. wrote the manuscript; and H.H. conceived the project.

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Correspondence to Hans Häcker.

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Redecke, V., Wu, R., Zhou, J. et al. Hematopoietic progenitor cell lines with myeloid and lymphoid potential. Nat Methods 10, 795–803 (2013). https://doi.org/10.1038/nmeth.2510

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