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Reprogramming fibroblasts to express T-cell functions using cell extracts

Nature Biotechnology volume 20, pages 460–466 (2002)Cite this article

Abstract

We demonstrate here the functional reprogramming of a somatic cell using a nuclear and cytoplasmic extract derived from another somatic cell type. Reprogramming of 293T fibroblasts in an extract from primary human T cells or from a transformed T-cell line is evidenced by nuclear uptake and assembly of transcription factors, induction of activity of a chromatin remodeling complex, histone acetylation, and activation of lymphoid cell–specific genes. Reprogrammed cells express T cell–specific receptors and assemble the interleukin-2 receptor in response to T cell receptor–CD3 (TCR–CD3) complex stimulation. Reprogrammed primary skin fibroblasts also express T cell–specific antigens. After exposure to a neuronal precursor extract, 293T fibroblasts express a neurofilament protein and extend neurite-like outgrowths. In vitro reprogramming of differentiated somatic cells creates possibilities for producing isogenic replacement cells for therapeutic applications.

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Figure 1: Import and chromatin binding of transcriptional activators of the IL-2 gene in 293T nuclei exposed to STE.
Figure 2: Chromatin remodeling and activation of the IL2 gene in STE.
Figure 3: 293T fibroblasts reprogrammed in Jurkat-TAg extract express hematopoietic cell-specific genes.
Figure 4: Fibroblasts reprogrammed in Jurkat extract exhibit hematopoietic cell markers.
Figure 5: Induction of a T cell–specific signaling pathway in reprogrammed cells.
Figure 6: Fibroblasts reprogrammed in NT2 extract express neurofilament protein NF200.

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Acknowledgements

We are grateful to T. Küntziger, A. Spurkland, and Y. Qi for discussion, primers, probes, and technical assistance. This work was supported by the Research Council of Norway and Nucleotech, LLC. P.C. is a fellow of the Norwegian Cancer Society.

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Authors and Affiliations

  1. Institute of Medical Biochemistry, P.O. Box 1112

    Anne-Mari Håkelien, Helga B. Landsverk & Philippe Collas

  2. Blindern, University of Oslo, Oslo, 0317, Norwayand

    Anne-Mari Håkelien, Helga B. Landsverk & Philippe Collas

  3. Institute for Nutrition Research, P.O. Box 1046

    Bjørn S. Skålhegg

  4. Blindern, University of Oslo, Oslo, 0317, Norway

    Bjørn S. Skålhegg

  5. Nucleotech LLC, 33 Riverside Avenue, Westport, 06880, CT

    Anne-Mari Håkelien, James M. Robl & Philippe Collas

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Correspondence to Philippe Collas.

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Competing interests

Part of this project was funded by Nucleotech, LLC, and A.-M.H. was employed by Nucleotech, LLC, during the completion of this work. The nuclear and cell reprogramming methods described here are the property of Nucleotech, LLC (PCT application no. PCT/US 01/47882).

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Håkelien, AM., Landsverk, H., Robl, J. et al. Reprogramming fibroblasts to express T-cell functions using cell extracts. Nat Biotechnol 20, 460–466 (2002). https://doi.org/10.1038/nbt0502-460

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