Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

Direct conversion of human fibroblasts to multilineage blood progenitors

Nature volume 468, pages 521–526 (2010)Cite this article

Subjects

An Author Correction to this article was published on 24 July 2018

This article has been updated

Abstract

As is the case for embryo-derived stem cells, application of reprogrammed human induced pluripotent stem cells is limited by our understanding of lineage specification. Here we demonstrate the ability to generate progenitors and mature cells of the haematopoietic fate directly from human dermal fibroblasts without establishing pluripotency. Ectopic expression of OCT4 (also called POU5F1)-activated haematopoietic transcription factors, together with specific cytokine treatment, allowed generation of cells expressing the pan-leukocyte marker CD45. These unique fibroblast-derived cells gave rise to granulocytic, monocytic, megakaryocytic and erythroid lineages, and demonstrated in vivo engraftment capacity. We note that adult haematopoietic programs are activated, consistent with bypassing the pluripotent state to generate blood fate: this is distinct from haematopoiesis involving pluripotent stem cells, where embryonic programs are activated. These findings demonstrate restoration of multipotency from human fibroblasts, and suggest an alternative approach to cellular reprogramming for autologous cell-replacement therapies that avoids complications associated with the use of human pluripotent stem cells.

This is a preview of subscription content, access via your institution

Access options

Rent or buy this article

Prices vary by article type

from$1.95

to$39.95

Learn more

Prices may be subject to local taxes which are calculated during checkout

Figure 1: OCT4 transduced human fibroblasts give rise to CD45+ve colonies.
Figure 2: OCT4 transduced dermal fibroblasts bypass the pluripotent state.
Figure 3: In vitro generation of myeloid lineages from CD45+ Fibs
Figure 4: In vivo capacity of CD45+FibsOCT4.
Figure 5: EPO treated CD45+FibsOCT4 generate erythroid and megakaryocytic progenitors.
Figure 6: OCT4 causes haematopoietic program activation in Fibs.

Similar content being viewed by others

Accession codes

Primary accessions

Gene Expression Omnibus

Data deposits

Data are available on the NCBI Gene Expression Omnibus (GEO) and are accessible through GEO Series accession number GSE24621.

Change history

  • 24 July 2018

    In this Article, there were duplicated empty lanes in Supplementary Figs. 2e and 3b. The corrected figures are presented in the Supplementary Information to the accompanying Amendment. The original Article has not been corrected.

References

  1. Jaenisch, R. & Young, R. Stem cells, the molecular circuitry of pluripotency and nuclear reprogramming. Cell 132, 567–582 (2008)

    CAS  PubMed  PubMed Central  Google Scholar 

  2. Chan, E. M. et al. Live cell imaging distinguishes bona fide human iPS cells from partially reprogrammed cells. Nature Biotechnol. 27, 1033–1037 (2009)

    CAS  Google Scholar 

  3. Lin, T. et al. A chemical platform for improved induction of human iPSCs. Nature Methods 6, 805–808 (2009)

    CAS  PubMed  PubMed Central  Google Scholar 

  4. Mikkelsen, T. S. et al. Dissecting direct reprogramming through integrative genomic analysis. Nature 454, 49–55 (2008)

    ADS  CAS  PubMed  PubMed Central  Google Scholar 

  5. Kanawaty, A. & Henderson, J. Genomic analysis of induced pluripotent stem (iPS) cells: routes to reprogramming. Bioessays 31, 134–138 (2009)

    PubMed  Google Scholar 

  6. Feng, R. et al. PU.1 and C/EBPα/β convert fibroblasts into macrophage-like cells. Proc. Natl Acad. Sci. USA 105, 6057–6062 (2008)

    ADS  CAS  PubMed  PubMed Central  Google Scholar 

  7. Vierbuchen, T. et al. Direct conversion of fibroblasts to functional neurons by defined factors. Nature 463, 1035–1041 (2010)

    ADS  CAS  PubMed  PubMed Central  Google Scholar 

  8. Ieda, M. et al. Direct reprogramming of fibroblasts into functional cardiomyocytes by defined factors. Cell 142, 375–386 (2010)

    CAS  PubMed  PubMed Central  Google Scholar 

  9. Kang, J., Shakya, A. & Tantin, D. Stem cells, stress, metabolism and cancer: a drama in two Octs. Trends Biochem. Sci. 34, 491–499 (2009)

    CAS  PubMed  Google Scholar 

  10. Brunner, C. et al. B cell-specific transgenic expression of Bcl2 rescues early B lymphopoiesis but not B cell responses in BOB.1/OBF.1-deficient mice. J. Exp. Med. 197, 1205–1211 (2003)

    CAS  PubMed  PubMed Central  Google Scholar 

  11. Emslie, D. et al. Oct2 enhances antibody-secreting cell differentiation through regulation of IL-5 receptor α chain expression on activated B cells. J. Exp. Med. 205, 409–421 (2008)

    CAS  PubMed  PubMed Central  Google Scholar 

  12. Pfisterer, P. et al. CRISP-3, a protein with homology to plant defense proteins, is expressed in mouse B cells under the control of Oct2. Mol. Cell. Biol. 16, 6160–6168 (1996)

    CAS  PubMed  PubMed Central  Google Scholar 

  13. Takahashi, K. et al. Induction of pluripotent stem cells from adult human fibroblasts by defined factors. Cell 131, 861–872 (2007)

    CAS  PubMed  Google Scholar 

  14. Takahashi, K. & Yamanaka, S. Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors. Cell 126, 663–676 (2006)

    CAS  PubMed  Google Scholar 

  15. Yu, J. et al. Induced pluripotent stem cell lines derived from human somatic cells. Science 318, 1917–1920 (2007)

    ADS  CAS  PubMed  Google Scholar 

  16. Hassan, H. T. & Zander, A. Stem cell factor as a survival and growth factor in human normal and malignant hematopoiesis. Acta Haematol. 95, 257–262 (1996)

    CAS  PubMed  Google Scholar 

  17. Lyman, S. D. et al. Molecular cloning of a ligand for the flt3/flk-2 tyrosine kinase receptor: a proliferative factor for primitive hematopoietic cells. Cell 75, 1157–1167 (1993)

    CAS  PubMed  Google Scholar 

  18. Kim, J. B. et al. Direct reprogramming of human neural stem cells by OCT4. Nature 461, 649–653 (2009)

    ADS  CAS  PubMed  Google Scholar 

  19. Lebofsky, R. & Walter, J. C. New Myc-anisms for DNA replication and tumorigenesis? Cancer Cell 12, 102–103 (2007)

    CAS  PubMed  Google Scholar 

  20. Silverstein, S. C., Steinman, R. M. & Cohn, Z. A. Endocytosis. Annu. Rev. Biochem. 46, 669–722 (1977)

    CAS  PubMed  Google Scholar 

  21. Hope, K. J., Jin, L. & Dick, J. E. Acute myeloid leukemia originates from a hierarchy of leukemic stem cell classes that differ in self-renewal capacity. Nature Immunol. 5, 738–743 (2004)

    CAS  Google Scholar 

  22. Roy, N. S. et al. Functional engraftment of human ES cell-derived dopaminergic neurons enriched by coculture with telomerase-immortalized midbrain astrocytes. Nature Med. 12, 1259–1268 (2006)

    CAS  PubMed  Google Scholar 

  23. Amariglio, N. et al. Donor-derived brain tumor following neural stem cell transplantation in an ataxia telangiectasia patient. PLoS Med. 6, e1000029 (2009)

    PubMed  PubMed Central  Google Scholar 

  24. Fried, W. Erythropoietin and erythropoiesis. Exp. Hematol. 37, 1007–1015 (2009)

    CAS  PubMed  Google Scholar 

  25. Perlingeiro, R. C., Kyba, M. & Daley, G. Q. Clonal analysis of differentiating embryonic stem cells reveals a hematopoietic progenitor with primitive erythroid and adult lymphoid-myeloid potential. Development 128, 4597–4604 (2001)

    CAS  PubMed  Google Scholar 

  26. Debili, N. et al. Characterization of a bipotent erythro-megakaryocytic progenitor in human bone marrow. Blood 88, 1284–1296 (1996)

    CAS  PubMed  Google Scholar 

  27. Klimchenko, O. et al. A common bipotent progenitor generates the erythroid and megakaryocyte lineages in embryonic stem cell-derived primitive hematopoiesis. Blood 114, 1506–1517 (2009)

    CAS  PubMed  Google Scholar 

  28. Strodtbeck, D. et al. Graft clonogenicity and intensity of pre-treatment: factors affecting outcome of autologous peripheral hematopoietic cell transplantation in patients with acute myeloid leukemia in first remission. Bone Marrow Transplant. 36, 1083–1088 (2005)

    CAS  PubMed  Google Scholar 

  29. Orkin, S. H. & Zon, L. I. Hematopoiesis and stem cells: plasticity versus developmental heterogeneity. Nature Immunol. 3, 323–328 (2002)

    CAS  Google Scholar 

  30. Shivdasani, R. A., Mayer, E. L. & Orkin, S. H. Absence of blood formation in mice lacking the T-cell leukaemia oncoprotein tal-1/SCL. Nature 373, 432–434 (1995)

    ADS  CAS  PubMed  Google Scholar 

  31. Ichikawa, M., Asai, T., Chiba, S., Kurokawa, M. & Ogawa, S. Runx1/AML-1 ranks as a master regulator of adult hematopoiesis. Cell Cycle 3, 722–724 (2004)

    CAS  PubMed  Google Scholar 

  32. Friedman, A. D. Transcriptional control of granulocyte and monocyte development. Oncogene 26, 6816–6828 (2007)

    CAS  PubMed  Google Scholar 

  33. Koschmieder, S., Rosenbauer, F., Steidl, U., Owens, B. M. & Tenen, D. G. Role of transcription factors C/EBPα and PU.1 in normal hematopoiesis and leukemia. Int. J. Hematol. 81, 368–377 (2005)

    CAS  PubMed  Google Scholar 

  34. Ng, E. S. et al. The primitive streak gene Mixl1 is required for efficient haematopoiesis and BMP4-induced ventral mesoderm patterning in differentiating ES cells. Development 132, 873–884 (2005)

    CAS  PubMed  Google Scholar 

  35. Tsai, F. Y. et al. An early haematopoietic defect in mice lacking the transcription factor GATA-2. Nature 371, 221–226 (1994)

    ADS  CAS  PubMed  Google Scholar 

  36. Vijayaragavan, K. et al. Noncanonical Wnt signaling orchestrates early developmental events toward hematopoietic cell fate from human embryonic stem cells. Cell Stem Cell 4, 248–262 (2009)

    CAS  PubMed  PubMed Central  Google Scholar 

  37. Boyer, L. A. et al. Core transcriptional regulatory circuitry in human embryonic stem cells. Cell 122, 947–956 (2005)

    CAS  PubMed  PubMed Central  Google Scholar 

  38. Kistler, B., Pfisterer, P. & Wirth, T. Lymphoid- and myeloid-specific activity of the PU.1 promoter is determined by the combinatorial action of octamer and ets transcription factors. Oncogene 11, 1095–1106 (1995)

    CAS  PubMed  Google Scholar 

  39. Rodda, D. J. et al. Transcriptional regulation of nanog by OCT4 and SOX2. J. Biol. Chem. 280, 24731–24737 (2005)

    CAS  PubMed  Google Scholar 

  40. Sridharan, R. et al. Role of the murine reprogramming factors in the induction of pluripotency. Cell 136, 364–377 (2009)

    CAS  PubMed  PubMed Central  Google Scholar 

  41. Ghozi, M. C., Bernstein, Y., Negreanu, V., Levanon, D. & Groner, Y. Expression of the human acute myeloid leukemia gene AML1 is regulated by two promoter regions. Proc. Natl Acad. Sci. USA 93, 1935–1940 (1996)

    ADS  CAS  PubMed  PubMed Central  Google Scholar 

  42. Chang, K. H. et al. Definitive-like erythroid cells derived from human embryonic stem cells coexpress high levels of embryonic and fetal globins with little or no adult globin. Blood 108, 1515–1523 (2006)

    CAS  PubMed  PubMed Central  Google Scholar 

  43. Kwon, U. K., Yen, P. H., Collins, T. & Wells, R. A. Differential lineage-specific regulation of murine CD45 transcription by Oct-1 and PU.1. Biochem. Biophys. Res. Commun. 344, 146–154 (2006)

    CAS  PubMed  Google Scholar 

  44. Feugier, P. et al. Hematologic recovery after autologous PBPC transplantation: importance of the number of postthaw CD34+ cells. Transfusion 43, 878–884 (2003)

    PubMed  Google Scholar 

  45. Rampalli, S. et al. p38 MAPK signaling regulates recruitment of Ash2L-containing methyltransferase complexes to specific genes during differentiation. Nature Struct. Mol. Biol. 14, 1150–1156 (2007)

    CAS  Google Scholar 

  46. Oshima, A. et al. Cloning, sequencing, and expression of cDNA for human β-glucuronidase. Proc. Natl Acad. Sci. USA 84, 685–689 (1987)

    ADS  CAS  PubMed  PubMed Central  Google Scholar 

  47. Li, C. & Wong, W. H. Model-based analysis of oligonucleotide arrays: expression index computation and outlier detection. Proc. Natl Acad. Sci. USA 98, 31–36 (2001)

    ADS  CAS  PubMed  MATH  Google Scholar 

Download references

Acknowledgements

This work was supported by grants to M.B. from the Canadian Institute of Health Research (CIHR), the Canadian Cancer Society Research Institute (CCS-RI), the StemCell Network and the Ontario Ministry of Research Innovation (MRI). M.B. is supported by the Canadian Chair Program and holds the Canada Research Chair in human stem cell biology. E.S. is supported by Ministry of Research and Innovation (MRI) and MITACS fellowships, R.M.R is supported by a CCS-RI fellowship and R.M. is supported by an Ontario Graduate Scholarship (OGS). We thank T. Werbowetski-Ogilvie for her help.

Author information

Authors and Affiliations

  1. Stem Cell and Cancer Research Institute, McMaster University, Hamilton, Ontario, Canada L8N 3Z5

    Eva Szabo, Shravanti Rampalli, Ruth M. Risueño, Angelique Schnerch, Ryan Mitchell, Aline Fiebig-Comyn, Marilyne Levadoux-Martin & Mickie Bhatia

  2. Department of Biochemistry and Biomedical Sciences, McMaster University, Hamilton, Ontario, Canada L8N 3Z5

    Angelique Schnerch, Ryan Mitchell & Mickie Bhatia

Authors
  1. Eva Szabo
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Shravanti Rampalli
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Ruth M. Risueño
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Angelique Schnerch
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Ryan Mitchell
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Aline Fiebig-Comyn
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Marilyne Levadoux-Martin
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Mickie Bhatia
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

All authors contributed to the acquisition, analysis and interpretation of the data; E.S., S.R., R.M.R. and M.B. initiated and designed the study; A.S. performed Affymetrix analyses; R.M.R. performed in vivo analyses; E.S., S.R., R.M.R. and M.B. wrote the paper.

Corresponding author

Correspondence to Mickie Bhatia.

Ethics declarations

Competing interests

The authors declare no competing financial interests.

Supplementary information

Supplementary Tables

This file contains Supplementary Tables 1 - 4. (PDF 897 kb)

Supplementary Figures

This file contains Supplementary Figures 1-18 with legends and additional references. (PDF 19780 kb)

PowerPoint slides

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Szabo, E., Rampalli, S., Risueño, R. et al. Direct conversion of human fibroblasts to multilineage blood progenitors. Nature 468, 521–526 (2010). https://doi.org/10.1038/nature09591

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/nature09591

This article is cited by

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing