Lineage conversion of one somatic cell type to another is an attractive approach for generating specific human cell types. Lineage conversion can be direct, in the absence of proliferation and multipotent progenitor generation, or indirect, by the generation of expandable multipotent progenitor states. We report the development of a reprogramming methodology in which cells transition through a plastic intermediate state, induced by brief exposure to reprogramming factors, followed by differentiation. We use this approach to convert human fibroblasts to mesodermal progenitor cells, including by non-integrative approaches. These progenitor cells demonstrated bipotent differentiation potential and could generate endothelial and smooth muscle lineages. Differentiated endothelial cells exhibited neo-angiogenesis and anastomosis in vivo. This methodology for indirect lineage conversion to angioblast-like cells adds to the armamentarium of reprogramming approaches aimed at the study and treatment of ischemic pathologies.
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We are thankful to Y. Zheng for his expertise and assistance with sorting procedures, C. Maiza for his expertise and assistance with in vivo procedures and M. Schwarz for administrative support. L.K. was partially supported by the California Institute for Regenerative Medicine. E.N. was partially supported by an F.M. Kirby Foundation postdoctoral fellowship. A.M.D. was supported by the Helmsley Foundation. L.C.L., R.D.T., F.S.B. and J.F.L. are supported by the California Institute for Regenerative Medicine (CL1-00502, RT1-01108, TR1-01250, RN2-00931), US National Institutes of Health (R33MH087925), US National Institutes of Health/National Institute Child Health and Human Development K12 Career Development Award (L.C.L.), Hartwell Foundation (L.C.L., R.D.T., F.S.B.), Millipore Foundation (J.F.L.) and Esther O'Keefe Foundation (J.F.L.). Work in the laboratory of F.H.G. was supported by the JPB Foundation, G. Harold and Leila Y. Mathers Charitable Foundation and Ellison Medical Foundation. Work in the laboratory of J.C.I.B. was supported by grants from Fundacion Cellex, the G. Harold and Leila Y. Mathers Charitable Foundation, the Leona M. and Harry B. Helmsley Charitable Trust, Sanofi, Ministerio de Economia y Competitividad (PLE2009-0100), Instituto de Salud Carlos III (ISCIII), Terapia Celular (TerCel) (RD06/0010/0016) and Fondo Europeo de Desarrollo Regional (FEDER).
The authors declare no competing financial interests.
Supplementary Figures 1–12 and Supplementary Tables 1 and 3 (PDF 23185 kb)
Specific mRNA fold change of the qPCR data summarized in Figures 1–3. (XLSX 54 kb)
Detailed mRNA and methylation array data including differentially regulated gene lists. (XLSX 22235 kb)
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Kurian, L., Sancho-Martinez, I., Nivet, E. et al. Conversion of human fibroblasts to angioblast-like progenitor cells. Nat Methods 10, 77–83 (2013). https://doi.org/10.1038/nmeth.2255
Generation of keratinocyte stem‐like cells from human fibroblasts via a direct reprogramming approach
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