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Culturing Keratinocytes on Biomimetic Substrates Facilitates Improved Epidermal Assembly In Vitro
Eve Hunter-Featherstone1, Natalie Young1, Kathryn Chamberlain1
1Department of Biosciences, Durham University, Durham DH1 3LE, UK.
Cells
|June 2, 2021
Summary
Researchers found that culturing skin cells on stiff plastic before 3D models alters their behavior. Using biomimetic hydrogels improved cell organization and 3D skin model development, highlighting the importance of mechanical cues in skin research.
Area of Science:
- Cell Biology
- Biomaterials Science
- Tissue Engineering
Background:
- Mechanotransduction enables cells to sense and respond to mechanical stimuli.
- Epidermal keratinocytes' behavior (proliferation, migration, differentiation) is influenced by mechanical cues.
- Standard in vitro cell culture uses stiff plastic, unlike the in vivo skin environment, potentially impacting epidermal models.
Purpose of the Study:
- To investigate the impact of pre-culture substrate stiffness on epidermal keratinocyte behavior.
- To compare keratinocytes cultured on tissue culture plastic versus biomimetic hydrogels with varying stiffness.
- To assess the influence of mechanical conditioning on the development of three-dimensional (3D) epidermal models.
Main Methods:
- Primary keratinocytes were cultured on tissue culture plastic and 4, 8, and 50 kPa biomimetic hydrogels.
- Evaluated changes in cellular architecture, cell density, nuclear biomechanics, and mechanoprotein expression (including Linker of Nucleoskeleton and Cytoskeleton complex constituents).
- Assessed the thickness and organization of 3D epidermal models derived from pre-conditioned keratinocytes.
Main Results:
- Keratinocytes cultured on biomimetic hydrogels showed significant alterations in cellular architecture, density, nuclear biomechanics, and mechanoprotein expression compared to those on plastic.
- Mechanical conditioning on 50 kPa hydrogels notably enhanced the thickness and organization of 3D epidermal models.
- Specific Linker of Nucleoskeleton and Cytoskeleton (LINC) complex constituents were identified as mechanoproteins affected by substrate stiffness.
Conclusions:
- Extracellular matrix mechanics profoundly influence keratinocyte cell biology.
- Pre-culturing keratinocytes on physiologically relevant, softer biomimetic hydrogels improves 3D epidermal model formation.
- Harnessing mechanical cues in skin research is crucial for developing accurate and physiologically relevant skin models.
