Direct conversion of human fibroblasts to multilineage blood progenitors

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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.

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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.

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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.

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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

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.

Correspondence to Mickie Bhatia.

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The authors declare no competing financial interests.

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

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