spacer silicone sheet  (Silex Silicones Ltd)

Journal: ACS Biomaterials Science & Engineering

Article Title: Modular Multiwell Viscoelastic Hydrogel Platform for Two- and Three-Dimensional Cell Culture Applications

doi: 10.1021/acsbiomaterials.4c00312

Figure Lengend Snippet: High-throughput fabrication approach to a 96-well hyaluronic acid hydrogel array. (a) Two modes of cross-linking were utilized in this work (covalent and supramolecular), both through the modification of hyaluronic acid (HA). Norbornene-modified HA (NorHA) is covalently cross-linked with dithiothreitol (DTT) through a light-mediated thiol–ene click reaction to form elastic hydrogels. Viscoelastic hydrogels additionally incorporate β-cyclodextrin-adamantane host–guest interactions, where a thiolated adamantane peptide is tethered to the NorHA through the same thiol–ene click reaction. (b) All components of the elastic or viscoelastic hydrogel networks come together as depicted in the schematic, where hydrogels can also be modified with thiolated RGD peptide to enable the culture of adhesive cells. (c) 96-well HA hydrogel arrays for 2D cell culture are fabricated through the following steps: (1) functionalization of glass piece with free thiols and application of silicone spacer sheet onto glass; (2) application of distinct hydrogel precursor solutions into cutouts in the silicone sheet; (3) flattening of hydrogel precursor solutions with SigmaCote-treated glass piece and curing of hydrogels by UV light; and (4) removal of top glass piece and silicone sheet and application of bottomless 96-well plate.

Article Snippet: For 2D cell culture, silicone spacer sheets of 0.5 mm thickness (Grace BioLabs, Seattle, WA) were laser cut to the 96-well geometry with a diameter of 0.5 mm less than the wells to enable some margin for improper alignment ( Figure S6 ).

Techniques: High Throughput Screening Assay, Modification, Adhesive, Cell Culture