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Cited by 114Cellular Mechanics and InteractionsCell Adhesion Molecules ResearchSkin and Cellular Biology Research

Expression of the differentiated phenotype by epithelial cells in vitro is regulated by both biochemistry and mechanics of the substratum

Opas, Michal·1989·Developmental Biology
DOI 10.1016/s0012-1606(89)80001-6
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Abstract

In this paper I sought to determine how the expression of differentiated traits of chick retinal pigmented epithelial (RPE) cells in vitro can be modulated by varying both the biochemical and the spatial complexity, and the mechanical properties, of the growth substratum. I have used glass derivatized with proteins of a basement membrane extract (nondeformable, two-dimensional substratum) and gels of reconstituted basement membrane extract (viscoelastic, three-dimensional substratum). These two biochemically similar substrata were compared to an inert substratum (untreated glass) and to the native basement membrane of the RPE, i.e., Bruch's Membrane. With immunofluorescence microscopy, I have shown that RPE cells, given space, will spread on their native basement membrane and form stress fibres and focal contacts, analogous to the stress fibres and integrin-, talin-, and vinculin-containing focal contacts of the cells grown on glass. Therefore, the stress fibres and focal contacts present in cultured cells are not artifacts of growth in vitro, but are a natural cellular response to the nondeformability of commonly used tissue culture substrata. The proteins of the basement membrane promote expression of some of the differentiated traits by RPE cells in vitro: however, the fully differentiated phenotype is expressed by RPE cells only when their spreading is prevented by low resilience of a substratum. Basement membrane gels generally are not resilient enough to support RPE cell spreading; however, the cells spread and form stress fibres, and integrin-, talin-, and vinculin-containing focal contacts when they are presented with areas of the gel which locally acquired higher resilience. The extent of cell spreading is determined by the deformability of substratum, hence elastic forces operating within the substratum determine the maximal cell traction allowable and, indirectly, the cytoarchitecture. Therefore, in addition to biochemical composition, the mechanical properties of substrata play important role in regulation of expression of the differentiated phenotype of cells in vitro and, possibly, in vivo.

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  • Polarity and differentiation of RPE cultures are influenced by the source of the cells, plating density, and the substrate on which the cells are grown.

    “Polarity and differentiation are not only influenced by the source of the cells (transformed, primary, stem cell-derived), but also by plating density (Adijanto and Philp, 2014; Toops et al., 2014), and the substrate on which the cells are grown (Opas, 1989).”

    From The cell biology of the retinal pigment epithelium (2020) · cites this paper as Opas, 1989

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Authors
Opas, M.
Venue
Developmental Biology
Publisher
Elsevier BV
Published
1989-02
Volume
131
Issue
2
Pages
281-293
ISSN
0012-1606 (ISSN-L)
DOI
10.1016/s0012-1606(89)80001-6
OpenAlex
W2088703323
Cited by
114
References
52
Citations by year
2025: 1 · 2020: 1 · 2017: 1 · 2016: 2 · 2013: 2 · 2012: 1
FWCI
3.97
Type
journal article
Language
en
Open access
closed
Licence
www.elsevier.com/tdm/userlicense/1.0/ (tdm)
Locations
  • Developmental Biology (publishedVersion)
  • PubMed (publishedVersion)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular Mechanics and Interactions
  • Health Sciences › Medicine › Immunology and Allergy › Cell Adhesion Molecules Research
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Skin and Cellular Biology Research
Keywords
retinal pigment epithelium, cell differentiation, substrate mechanics, substrate stiffness, basement membrane, Bruch's membrane, stress fibers, focal adhesions, integrins, talin, vinculin, cell spreading, viscoelastic gels, cell traction forces, chick embryo
MeSH terms
Animals, Antibodies, Antigens, Surface, Basement Membrane, Cell Adhesion, Cell Differentiation, Cell Division, Cells, Cultured, Chick Embryo, Cytoskeletal Proteins, Cytoskeleton, Fibronectins, Fluorescent Antibody Technique, Gene Expression Regulation, Glass, Membrane Glycoproteins, Muscle Proteins, Phenotype, Pigment Epithelium of Eye, Integrins, Vinculin, Talin

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