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. 2022 Feb;22(3):e2100116.
doi: 10.1002/pmic.202100116. Epub 2021 Nov 2.

Passages in culture and stimulation conditions influence protein expression of primary fibroblasts

Solange Vivier  1 , Aurélien Chepy  1   2 , Fabrice Bray  3 , Thomas Guerrier  1   4 , Silvia Speca  1 , Stéphanie Flament  3 , Manel Jendoubi  1 , Maïté Balden  1   4 , Christian Rolando  3 , Eric Hachulla  1   2 , David Launay  1   2 , Sylvain Dubucquoi  1   4 , Vincent Sobanski  1   2   5
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Passages in culture and stimulation conditions influence protein expression of primary fibroblasts

Solange Vivier et al. Proteomics. 2022 Feb.

Abstract

Fibroblasts (Fb) are key effector cells in systemic sclerosis (SSc). Fb stimulation with transforming growth factor beta 1 (TGF-β1) is considered as a positive control in studies assessing fibrogenesis. The lack of standardization of TGF-β1 stimulation might be responsible for discrepancies in experiments performed in different conditions. Using quantitative proteomics analysis, we evaluated the impact of changes in experimental conditions on proteomic profiles of primary Fb. Principal component analysis (PCA) identified several groups of differentially expressed proteins influenced by cell passage, culture medium, and both concentration and duration of exposure to TGF-β1 stimulation. Bioinformatics analysis revealed that late passages expressed proteins involved in senescence. TGF-β1 concentration and time of stimulation were correlated with the expression of proteins involved in the fibrogenesis and inflammatory processes. These data underline the need for standardization of culture conditions to allow inter-data comparisons in future in vitro studies, especially when using "omics" approaches.

Keywords: TGF-β1; fibrosis; primary fibroblasts; proteomics; systemic sclerosis.

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References

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