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Substrate softness promotes terminal differentiation of human keratinocytes without altering their ability to proliferate back into a rigid environment

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Abstract

Substrate stiffness is a key regulator of cell behavior. To investigate how mechanical properties of cell microenvironment affect the human keratinocyte, primary cells were seeded on polyacrylamide hydrogels of different compliances (soft: 4 kPa, medium: 14 kPa, rigid: 45 kPa) in comparison with glass coverslip (> GPa). Keratinocyte spreading and proliferation were strongly decreased on the softest hydrogel, while no significant difference was observed between medium, rigid hydrogels and glass coverslip, and cells’ viability was comparable in all conditions after 72 h. We then performed a RNA-seq to compare the transcriptomes from keratinocytes cultured for 72 h on the softest hydrogel or on coverslips. The cells on the soft hydrogel showed a strong increase in the expression of late differentiation marker genes from the epidermal differentiation complex (1q21) and the antioxidant machinery. In parallel, these cells displayed a significant loss of expression of the matrix receptors (integrin α6 and β1) and the EGF receptor. However, when these cells were replated on a plastic culture plate (> GPa), they were able to re-engage the proliferation machinery with a strong colony-formation efficiency. Overall, using low-calcium differentiation monolayers at confluence, the lesser the rigidity, the stronger the markers of late differentiation are expressed, while the inverse is observed regarding the markers of early differentiation. In conclusion, below a certain rigidity, human keratinocytes undergo genome reprogramming indicating terminal differentiation that can switch back to proliferation in contact with a stiffer environment.

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Acknowledgements

This work is supported by Isispharma France. Choua Ya is a recipient of a PhD grant from the French National Association of Research and Technology (ANRT). Mariana Carrancá is supported by a PhD grant from the Région Auvergne Rhône Alpes.

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Authors and Affiliations

  1. CNRS/UCBL, University Lyon 1, UMR 5305, Laboratory of Tissue Biology and Therapeutic Engineering, LBTI, IBCP, 7, passage du Vercors, 69367, Lyon cedex 7, France

    Choua Ya, Mariana Carrancá, Dominique Sigaudo-Roussel, Bérengère Fromy & Romain Debret

  2. Isispharma, 29 Rue Maurice Flandin, 69003, Lyon, France

    Choua Ya

  3. Alpol Cosmétique, 140 Rue Pasteur, 01500, Château-Gaillard, France

    Philippe Faure

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  1. Choua Ya
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Correspondence to Romain Debret.

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Ya, C., Carrancá, M., Sigaudo-Roussel, D. et al. Substrate softness promotes terminal differentiation of human keratinocytes without altering their ability to proliferate back into a rigid environment. Arch Dermatol Res 311, 741–751 (2019). https://doi.org/10.1007/s00403-019-01962-5

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