Live dead kits for mammalian cells

Cell viability assays were carried out at days 1 and 7 using Live/Dead kits for mammalian cells (Life Technologies). After washing all samples in warm PBS, a mixed solution of ethidium homodimer-1 and calcein was prepared in a ratio of 4:1 and 300 μL of this solution and incubated for 15 min. After this period, samples were washed in PBS five more times, before being left in PBS for microscope imaging.

The sodium salt of alginic acid (Alg, viscosity = 20 – 40 cPs) was supplied by Sigma Aldrich (St. Louis, MO). Acros Organics supplied calcium chloride and glycerol. αMEM and DMEM were supplied by GIBCO. Fetal bovine serum (FBS) was purchased from Thermo Scientific. Penicillin/streptomycin (P/S), trypsin EDTA and Amphotericin B were obtained from Corning Cellgro. Live/Dead kits for mammalian cells were supplied by Life Technologies. Glycolide and D,L-lactide were purchased from Polysciences (Warrington, PA). Lysine triisocyanate-poly(ethylene glycol) (LTI-PEG) prepolymer was supplied by Medtronic, Inc, and hexamethylene diisocyanate trimer (HDIt) was supplied by Bayer Material Science. Iron acetylacetonate (FeAA) was supplied by Sigma-Aldrich. ε-caprolactone was dried over anhydrous MgSO₄, and all other materials were used as received.