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Borosilicate glass surpasses polystyrene in optical quality; however, it is less frequently used for cell culture due to poor protein and cell adhesion. To overcome this impasse, the surface of glass coverslips requires functionalization to enable facile covalent attachment of proteins to promote cell attachment and differentiation. Herein, a novel approach is presented to covalently attach proteins to glass by depositing a thin layer of radical-rich carbon film using a plasma polymerization process. The surface chemistry of these plasma-activated coatings can be controlled by varying the gas composition used during the deposition. Mass spectrometry reveals different protein profiles attached to functionalized glass coverslips when they are exposed to cell culture media. Mouse embryonic stem cell adhesion and subsequent differentiation into neural lineage on plasma-treated coverslips are significantly enhanced compared to bare coverslips. Importantly, the coatings are in the nanometer range, preserve the optical properties of the glass coverslips for imaging, and remain stable for at least 4 weeks in simulated body fluid. These results demonstrate the utility of covalently attaching proteins to glass for enhanced cell attachment and stem cell differentiation and provide a promising technique to achieve better outcomes in cell culture in a range of biomedical applications.
Pubmed ID: 40213000
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View all literature mentionsA software package and server used to identify and characterize proteins from primary sequence databases using mass spectrometry data. Mascot integrates peptide mass fingerprinting, sequence querying, and MS/MS ion searching in order to search for proteins in databases like SwissProt, NCBInr, EMBL EST divisions, contaminants, and cRAP. If a license is purchased, users may: search data sets that exceed the 1200 spectrum limit of the free version; set up automated, high throughput work; add and edit proteins and quantification methods; and search a preferred collection of sequence databases. The software package works with instruments from AB Sciex, Agilent, Bruker, Jeol, Shimadzu, Thermo Scientific, and Waters.
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