Substrate Elastic Modulus Regulates the Morphology, Focal Adhesions, and α-Smooth Muscle Actin Expression of Retinal Müller Cells

• Published in: Investigative ophthalmology & visual science 2015-09, Vol.56 (10), p.5974-5982
• Main Authors: Bu, Shao-Chong, Kuijer, Roel, van der Worp, Roelofje J, van Putten, Sander M, Wouters, Olaf, Li, Xiao-Rong, Hooymans, Johanna M M, Los, Leonoor I
• Format: Article
• Language: English
• Subjects: Actins - metabolism; Cell Culture Techniques - methods; Cell Transdifferentiation - physiology; Cells, Cultured; Elastic Modulus - physiology; Ependymoglial Cells - drug effects; Ependymoglial Cells - metabolism; Ependymoglial Cells - physiology; Focal Adhesions - physiology; Humans; Immunohistochemistry; Transforming Growth Factor beta - pharmacology
• ISSN: 1552-5783; 1552-5783
• DOI: 10.1167/iovs.14-15969

The stiffness of the extracellular matrix has been shown to regulate cell adhesion, migration, and transdifferentiation in fibrotic processes. Retinal Müller cells have been shown to be mechanosensitive; they are involved in fibrotic vitreoretinal diseases. Since fibrosis increases the rigidity of the extracellular matrix, our aim was to develop an in vitro model for studying Müller cell morphology and differentiation state in relation to matrix stiffness. A spontaneously immortalized human Müller cell line (MIO-M1) was cultured on type I collagen-coated polyacrylamide gels with Young's moduli ranging from 2 to 92 kPa. Cell surface area, focal adhesion, and the expression and morphology of α-smooth muscle actin induced by transforming growth factor β (TGF-β [10 ng/mL for 48 hours]) were analyzed by immunocytology. The images were documented by using fluorescence microscopy and confocal scanning laser microscopy. MIO-M1 cells cultured on stiff substrates exhibited a significant increase in cell surface area, stress fiber, and mature focal adhesion formation. Furthermore, Müller cells treated with TGF-β1 and TGF-β2 and cultured on stiff substrates showed an increased incorporation of α-smooth muscle actin into stress fibers when compared to those grown on soft surfaces. Compliance of the surrounding matrix seems to influence the morphology and contraction of retinal Müller cells in fibrotic conditions. Development of an in vitro model simulating both the normally compliant retinal tissue and the rigid retinal fibrotic tissue helps fill the gap between the results of petri-dish cell culture with rigid surfaces and in vivo findings.