Fibronectin localization and fibrillization are affected by the presence of serum in culture media

Scientific Reports
Alessandro SianiNicola Tirelli

Abstract

In vitro models of fibrotic phenomena are often based on the fibroblast-myofibroblast transition as the contraction-triggering cellular event. There are, however, multiple sources of concern regarding the appropriateness of such models; a first and widely investigated issue is the often inappropriate nature of the interactions between mesenchymal cells and surrounding/underlying matrix/substrate. A second set of problems concerns the composition of the fluid phase, which includes both dispersed/dissolved paracrine messengers and matrix elements. In this study, we have focused on the effects that serum may generate. We have observed that A) serum causes high variability in the expression of typical markers of myofibroblast differentiation (ED-A fibronectin and α-Smooth Muscle Actin) upon treatment with TGF-β1; this is probably due to intrinsic variability of cytokine concentrations in different batches of serum. B) the fibrillization of endogenous fibronectin is partially hampered and its localization changed from ventral (on the substrate) to dorsal (upper surface); the latter morphology appears to be largely overlooked in literature, even though it may have a significant role in terms of mechanotransductive signaling. This qui...Continue Reading

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Citations

Jul 18, 2020·Inflammatory Bowel Diseases·Calen A SteinerPeter D R Higgins

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Methods Mentioned

BETA
ELISA
Atomic Force Microscopy
Fluorescence
AFM
confocal microscopy

Software Mentioned

MetaVue [UNK] Research Imaging
confocal
Elements

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