Macromolecular crowding regulates matrix composition and gene expression in human gingival fibroblast cultures.
Journal
Scientific reports
ISSN: 2045-2322
Titre abrégé: Sci Rep
Pays: England
ID NLM: 101563288
Informations de publication
Date de publication:
04 02 2023
04 02 2023
Historique:
received:
16
11
2022
accepted:
01
02
2023
pubmed:
5
2
2023
medline:
8
2
2023
entrez:
4
2
2023
Statut:
epublish
Résumé
Standard cell cultures are performed in aqueous media with a low macromolecule concentration compared to tissue microenvironment. In macromolecular crowding (MMC) experiments, synthetic polymeric crowders are added into cell culture media to better mimic macromolecule concentrations found in vivo. However, their effect on cultured cells is incompletely understood and appears context-dependent. Here we show using human gingival fibroblasts, a cell type associated with fast and scarless wound healing, that MMC (standard medium supplemented with Ficoll 70/400) potently modulates fibroblast phenotype and extracellular matrix (ECM) composition compared to standard culture media (nMMC) over time. MMC significantly reduced cell numbers, but increased accumulation of collagen I, cellular fibronectin, and tenascin C, while suppressing level of SPARC (Secreted Protein Acidic and Cysteine Rich). Out of the 75 wound healing and ECM related genes studied, MMC significantly modulated expression of 25 genes compared to nMMC condition. MMC also suppressed myofibroblast markers and promoted deposition of basement membrane molecules collagen IV, laminin 1, and expression of LAMB3 (Laminin Subunit Beta 3) gene. In cell-derived matrices produced by a novel decellularization protocol, the altered molecular composition of MMC matrices was replicated. Thus, MMC may improve cell culture models for research and provide novel approaches for regenerative therapy.
Identifiants
pubmed: 36739306
doi: 10.1038/s41598-023-29252-1
pii: 10.1038/s41598-023-29252-1
pmc: PMC9899282
doi:
Substances chimiques
Collagen Type I
0
Macromolecular Substances
0
Culture Media
0
Fibronectins
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
2047Subventions
Organisme : CIHR
ID : PJT-153223
Pays : Canada
Informations de copyright
© 2023. The Author(s).
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