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Use of a PTFE Micro-Bioreactor to Promote 3D Cell Rearrangement and Maintain High Plasticity in Epigenetically Erased Fibroblasts

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Abstract

Phenotype definition is driven by epigenetic mechanisms as well as directly influenced by the cell microenvironment and by biophysical signals deriving from the extracellular matrix. The possibility to interact with the epigenetic signature of an adult mature cell, reversing its differentiated state and inducing a short transient high plasticity window, was previously demonstrated. In parallel, in vitro studies have shown that 3D culture systems, mimicking cell native tissue, exert significant effects on cell behavior and functions. Here we report the production of “PTFE micro-bioreactors” for long-term culture of epigenetically derived high plasticity cells. The system promotes 3D cell rearrangement, global DNA demethylation and elevated transcription of pluripotency markers, that is dependent on WW domain containing transcription regulator 1 (TAZ) nuclear accumulation and SMAD family member 2 (SMAD2) co-shuttling. Our findings demonstrate that the use of 3D culture strategies greatly improves the induction and maintenance of a high plasticity state.

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Acknowledgements

This work was funded by Carraresi Foundation. The Laboratory of Biomedical Embryology is member of the COST Action CA16119 In vitro 3-D total cell guidance and fitness (CellFit), and the COST Action CM1406 Epigenetic Chemical Biology (EPICHEM). We thank Prof. E. Somigliana for kindly providing human skin biopsies.

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Author notes

  1. Georgia Pennarossa and Elena F. M. Manzoni contributed equally to the work.

Authors and Affiliations

  1. Laboratory of Biomedical Embryology, Department of Health, Animal Science and Food Safety, Università degli Studi di Milano, 20133, Milan, Italy

    Georgia Pennarossa, Elena F. M. Manzoni & Tiziana A. L. Brevini

  2. Department of Veterinary Medicine, University of Sassari, 07100, Sassari, Italy

    Sergio Ledda

  3. Department of Biotechnology and Life Sciences, Università degli Studi dell’Insubria, 21100, Varese, Italy

    Magda deEguileor

  4. Laboratory of Biomedical Embryology, Department of Agricultural and Environmental Sciences - Production, Landscape, Agroenergy, University of Milan, 20133, Milan, Italy

    Fulvio Gandolfi

  5. Unistem, Centre for Stem Cell Research, Universita’ degli Studi di Milano, 20133, Milan, Italy

    Fulvio Gandolfi & Tiziana A. L. Brevini

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  1. Georgia Pennarossa
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  2. Elena F. M. Manzoni
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  3. Sergio Ledda
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  4. Magda deEguileor
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  5. Fulvio Gandolfi
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  6. Tiziana A. L. Brevini
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Correspondence to Tiziana A. L. Brevini.

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Pennarossa, G., Manzoni, E.F.M., Ledda, S. et al. Use of a PTFE Micro-Bioreactor to Promote 3D Cell Rearrangement and Maintain High Plasticity in Epigenetically Erased Fibroblasts. Stem Cell Rev and Rep 15, 82–92 (2019). https://doi.org/10.1007/s12015-018-9862-5

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