Biomimetic dextran-based hydrogel layers for cell micropatterning over large areas using the FluidFM BOT technology
Langmuir, Just Accepted Manuscript
DOI: 10.1021/acs.langmuir.8b03249
Publication Date (Web): January 17, 2019
Copyright © 2019 American Chemical Society
Abstract
Micropatterning of living single cells and cell clusters over millimeter-centimeter scale areas is of high demand in the development cell-based biosensors. Micropatterning methodologies require both to have a suitable biomimetic support and printing technology. In this work we present the micropatterning of living mammalian cells on carboxymethyl dextran (CMD) hydrogel layers using the FluidFM BOT technology. In contrast to the ultrathin (few nanometers thick in dry state) CMD films generally used in label-free biosensor applications, we developed CMD layers with thicknesses of several tens of nanometers in order to provide support for the controlled adhesion of living cells. The fabrication method and detailed characterization of CMD layers are also described.The antifouling ability of the CMD surfaces is demonstrated by in situ optical waveguide lightmode spectroscopy (OWLS) measurements using serum modelling proteins with different electrostatic properties and molecular weights.
Cell micropatterning on the CMD surface was obtained by printing cell adhesion mediating cRGDfK peptide molecules (cyclo(Arg-Gly-Asp-D-Phe-Lys)) directly from aqueous solution using microchanneled cantilevers with subsequent incubation of the printed surfaces in living cell culture. Uniquely, we present cell patterns with different geometries (spot, line, grid arrays) covering both micrometer and millimeter-centimeter scale areas.
The adhered patterns were analyzed by phase contrast microscopy and the adhesion process on the patterns was real-time monitored by digital holographic microscopy, enabling to quantify the survival and migration of cells on the printed cRGDfK arrays.
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....Digital holographic microscopy (DHM) DHM was used to real-time monitor the adhesion of HeLa cells. Real-time holographic images were recorded by a HoloMonitor® M4 instrument (Phase Holographic Imaging PHI AB....
....The captured images were analyzed and formatted using the HoloStudio M4 software (Phase Holographic Imaging PHI AB)....
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