Thiols stabilize cobblestone morphology of cultured mesothelial cells

Abstract

Cellular thiols including GSH (glutathione) and L-Cys (L-cysteine) are essential for cell signalling, growth and differentiation. L-Cys is derived from the extracellular thiol pool and is the rate-limiting compound for intracellular GSH biosynthesis. The present study investigated the effect of thiol-supplemented medium on cell growth, phenotype and total GSH of cultured hPMCs (human peritoneal mesothelial cells). Cells were cultured in medium M199 supplemented with 2% serum, with 'plus' or without 'minus' L-Cys and compared with medium supplemented with either β-ME (β-mercaptoethanol) (0.25 mmol/l) or the receptor tyrosine kinase ligand EGF (epidermal growth factor, 100 ng/ml). β-ME produced a disproportionate increase in total GSH compared with L-Cys and other thiols tested [(procysteine (2-oxothiazolidine-4-carboxylic acid) or NAC (N-acetyl-L-cysteine)], while growth and morphology were identical. Cell behaviour of primary hPMCs is characterized by the transition of fibroblastoid to cobblestone morphology during early passage. L-Cys and β-ME promoted a rapid MET (mesenchymal-to-epithelial transition) within 3 days of culture, confirmed by the presence of cobblestone cells, intact organelles, abundant microvilli, primary cilia and cortical actin. In contrast, EGF produced a biphasic response consisting of delayed growth and retention of a fibroblastoid morphology. During a rapid log phase of growth, MET was accompanied by rapid catch-up growth. Thiols may stabilize the epithelial phenotype by engaging redox-sensitive receptors and transcription factors that modulate differentiation. These data may benefit researchers working on thiol-mediated cell differentiation and strategies to regenerate damage to serosal membranes.

Beta-Mecaptoethanol Suppresses Inflammation and Induces Adipogenic Differentiation in 3T3-F442A Murine Preadipocytes

“…As a thiol compound, BME is well known for its strong anti-oxidant power, in part through its effect to increase synthesis of thiol-containing redox couples [38]. In light of the link between impaired preadipocyte differentiation and oxidant stress associated with fat tissue inflammation, we asked whether BME could regulate adipogenic differentiation through modulation of the inflammatory pathways.…”

Modulation of mTOR and autophagy in hibernating hamster lung and the application of the potential mechanism to improve the recellularization process of decellularized lung scaffolds

“…Indeed, it has been previously shown by researchers that probably DPI and surprisingly NAC (N-acetyl-L-cysteine) might be inhibiting cell proliferation through different mechanism affecting cell cycle [29]. Indeed, it should not be neglected that there is literature pointing to the fact that active thiol chemistry is very important for cell growth and viability, affecting many signaling pathways [30,31]. Although the direct effects of -SH groups on recellularization is beyond the scope of this paper, the presence of higher free -SH groups through serotonin treatment might play a part in the positive outcome observed in serotonin treated recellularized lung.…”