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Culturing Keratinocytes on Biomimetic Substrates Facilitates Improved Epidermal Assembly In Vitro

Hunter-Featherstone, Eve; Young, Natalie; Chamberlain, Kathryn; Cubillas, Pablo; Hulette, Ben; Wei, Xingtao; Tiesman, Jay P.; Bascom, Charles C.; Benham, Adam M.; Goldberg, Martin W.; Saretzki, Gabriele; Karakesisoglou, Iakowos

Culturing Keratinocytes on Biomimetic Substrates Facilitates Improved Epidermal Assembly In Vitro Thumbnail


Authors

Eve Hunter-Featherstone

Kathryn Chamberlain

Pablo Cubillas

Ben Hulette

Xingtao Wei

Jay P. Tiesman

Charles C. Bascom

Gabriele Saretzki



Abstract

Mechanotransduction is defined as the ability of cells to sense mechanical stimuli from their surroundings and translate them into biochemical signals. Epidermal keratinocytes respond to mechanical cues by altering their proliferation, migration, and differentiation. In vitro cell culture, however, utilises tissue culture plastic, which is significantly stiffer than the in vivo environment. Current epidermal models fail to consider the effects of culturing keratinocytes on plastic prior to setting up three-dimensional cultures, so the impact of this non-physiological exposure on epidermal assembly is largely overlooked. In this study, primary keratinocytes cultured on plastic were compared with those grown on 4, 8, and 50 kPa stiff biomimetic hydrogels that have similar mechanical properties to skin. Our data show that keratinocytes cultured on biomimetic hydrogels exhibited major changes in cellular architecture, cell density, nuclear biomechanics, and mechanoprotein expression, such as specific Linker of Nucleoskeleton and Cytoskeleton (LINC) complex constituents. Mechanical conditioning of keratinocytes on 50 kPa biomimetic hydrogels improved the thickness and organisation of 3D epidermal models. In summary, the current study demonstrates that the effects of extracellular mechanics on keratinocyte cell biology are significant and therefore should be harnessed in skin research to ensure the successful production of physiologically relevant skin models

Citation

Hunter-Featherstone, E., Young, N., Chamberlain, K., Cubillas, P., Hulette, B., Wei, X., …Karakesisoglou, I. (2021). Culturing Keratinocytes on Biomimetic Substrates Facilitates Improved Epidermal Assembly In Vitro. Cells, 10(5), https://doi.org/10.3390/cells10051177

Journal Article Type Article
Acceptance Date May 9, 2021
Online Publication Date May 12, 2021
Publication Date 2021
Deposit Date Oct 11, 2021
Publicly Available Date Oct 11, 2021
Journal Cells
Electronic ISSN 2073-4409
Publisher MDPI
Peer Reviewed Peer Reviewed
Volume 10
Issue 5
DOI https://doi.org/10.3390/cells10051177

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Publisher Licence URL
http://creativecommons.org/licenses/by/4.0/

Copyright Statement
This article is an open access article
distributed under the terms and
conditions of the Creative Commons
Attribution (CC BY) license (https://
creativecommons.org/licenses/by/
4.0/).






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