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In in vitro live-cell imaging, itwould be beneficial to grow andassess human embryonic stem cell-derived retinal pigment epithelial(hESC-RPE) cells on thin, transparent, rigid surfaces such as coverglasses. In this study, we assessed how the silanization of glasswith 3-aminopropyltriethoxysilane (APTES), 3-(trimethoxysilyl)propylmethacrylate (MAPTMS), or polymer–ceramic material Ormocompaffects the surface properties, protein binding, and maturation ofhESC-RPE cells. The surface properties were studied by contact anglemeasurements, X-ray photoelectron spectroscopy (XPS), atomic forcemicroscopy (AFM), and a protein binding assay. The cell adherenceand proliferation were evaluated by culturing hESCRPE cells on collagenIV-coated untreated or silanized surfaces for 42 days. The Ormocomptreatment significantly increased the hydrophobicity and roughnessof glass surfaces compared to the APTES and MAPTMS treatments. TheXPS results indicated that the Ormocomp treatment changes the chemicalcomposition of the glass surface by increasing the carbon contentand the number of C–O/O bonds. The protein-bindingtest confirmed that the Ormocomp-treated surfaces bound more collagenIV than did APTES- or MAPTMS-treated surfaces. All of the silane treatmentsincreased the number of cells: after 42 days of culture, Ormocomphad 0.38, APTES had 0.16, MAPTMS had 0.19, and untreated glass hadonly 0.062, all presented as million cells cm–2.There were no differences in cell numbers compared to smoother torougher Ormocomp surfaces, suggesting that the surface chemistry and,more specifically, the collagen binding in combination with Ormocompare beneficial to hESC-RPE cell culture. This study clearly demonstratesthat Ormocomp treatment combined with collagen coating significantlyincreases hESC-RPE cell attachment compared to commonly used silanizingagents APTES and MAPTMS. Ormocomp silanization could thus enable theuse of microscopic live cell imaging methods for hESC-RPE cells. [ABSTRACT FROM AUTHOR]