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

Autor: Vivier S; Univ. Lille, Inserm, CHU Lille, U1286 - INFINITE - Institute for Translational Research in Inflammation, Lille, France., Chepy A; Univ. Lille, Inserm, CHU Lille, U1286 - INFINITE - Institute for Translational Research in Inflammation, Lille, France.; CHU Lille, Département de Médecine Interne et Immunologie Clinique, Centre de Référence des Maladies Auto-immunes Systémiques Rares du Nord et Nord-Ouest de France (CeRAINO), Lille, France., Bray F; Univ. Lille, CNRS, USR 3290 - MSAP - Miniaturisation pour la Synthèse, l'Analyse et la Protéomique, Lille, France., Guerrier T; Univ. Lille, Inserm, CHU Lille, U1286 - INFINITE - Institute for Translational Research in Inflammation, Lille, France.; Institut d'Immunologie, Centre de Biologie Pathologie, CHU Lille, Lille, France., Speca S; Univ. Lille, Inserm, CHU Lille, U1286 - INFINITE - Institute for Translational Research in Inflammation, Lille, France., Flament S; Univ. Lille, CNRS, USR 3290 - MSAP - Miniaturisation pour la Synthèse, l'Analyse et la Protéomique, Lille, France., Jendoubi M; Univ. Lille, Inserm, CHU Lille, U1286 - INFINITE - Institute for Translational Research in Inflammation, Lille, France., Balden M; Univ. Lille, Inserm, CHU Lille, U1286 - INFINITE - Institute for Translational Research in Inflammation, Lille, France.; Institut d'Immunologie, Centre de Biologie Pathologie, CHU Lille, Lille, France., Rolando C; Univ. Lille, CNRS, USR 3290 - MSAP - Miniaturisation pour la Synthèse, l'Analyse et la Protéomique, Lille, France., Hachulla E; Univ. Lille, Inserm, CHU Lille, U1286 - INFINITE - Institute for Translational Research in Inflammation, Lille, France.; CHU Lille, Département de Médecine Interne et Immunologie Clinique, Centre de Référence des Maladies Auto-immunes Systémiques Rares du Nord et Nord-Ouest de France (CeRAINO), Lille, France., Launay D; Univ. Lille, Inserm, CHU Lille, U1286 - INFINITE - Institute for Translational Research in Inflammation, Lille, France.; CHU Lille, Département de Médecine Interne et Immunologie Clinique, Centre de Référence des Maladies Auto-immunes Systémiques Rares du Nord et Nord-Ouest de France (CeRAINO), Lille, France., Dubucquoi S; Univ. Lille, Inserm, CHU Lille, U1286 - INFINITE - Institute for Translational Research in Inflammation, Lille, France.; Institut d'Immunologie, Centre de Biologie Pathologie, CHU Lille, Lille, France., Sobanski V; Univ. Lille, Inserm, CHU Lille, U1286 - INFINITE - Institute for Translational Research in Inflammation, Lille, France.; CHU Lille, Département de Médecine Interne et Immunologie Clinique, Centre de Référence des Maladies Auto-immunes Systémiques Rares du Nord et Nord-Ouest de France (CeRAINO), Lille, France.; Institut Universitaire de France (IUF), France.
Jazyk: angličtina
Zdroj: Proteomics [Proteomics] 2022 Feb; Vol. 22 (3), pp. e2100116. Date of Electronic Publication: 2021 Nov 02.
DOI: 10.1002/pmic.202100116
Abstrakt: 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.
(© 2021 Wiley-VCH GmbH.)
Databáze: MEDLINE
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