Low cell binding 96 well plates

L-aspartic acid (Sigma-Aldrich, UK), 1,4-diaminobutane (DAB) (Sigma-Aldrich, ≥99%), cysteamine (CYSE) (Sigma-Aldrich, UK), cystamine (CYS) (Sigma-Aldrich, UK), dimethylformamide (DMF) (VWR International, USA), dimethylsulfoxide (DMSO) (Sigma-Aldrich), o-phosphoric acid (VWR), imidazole (ACS reagent, ≥99%, Sigma-Aldrich), citric-acid*H₂O (ACS reagent, ≥99.9%, VWR), sodium chloride (99–100.5%, Sigma-Aldrich), phosphate buffer saline (PBS) (Tablet, Sigma), D,L-dithiotreitol (DTT) (Sigma), 5,5 dithio bis-(2-nitrobenzoic acid) (Sigma, ≥98%, USA), L-cystein (Sigma, ≥97%, USA), Humidified incubator (Nuaire, USA), 100 mm tissue culture dishes (Orange Scientific, Belgium), 48 well plates (Sigma-Aldrich, USA), low cell binding 96 well plates (Nunc, Denmark), Eagle’s Medium Alfa minimal essential medium (αMEM) (Gibco, USA), fetal bovine serum (FBS, Gibco, USA), L-glutamine (Gibco, USA), penicillin and streptomycin (Gibco, USA), L-ascorbic acid 2-phosphate (Sigma-Aldrich, USA), beta-glycerophosphate (Sigma-Aldrich, USA), dexamethasone (Sigma-Aldrich, USA), WST-1 [2-(4-Iodophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium] (Roche, Switzerland), Vybrant DiD (Molecular Probes, USA), 2-Amino-2-Methyl-1-Propanol buffer (Sigma-Aldrich, USA), Alkaline Phosphatase Yellow (pNPP) Liquid Substrate System (Sigma-Aldrich, USA).

To measure cell viability, gel discs with a diameter of 3 mm were prepared and placed in low cell binding 96 well plates (Nunc, Denmark). 20 000 PDLCs in 200 μl culture medium were seeded on each gel disc. After culturing for 1, 3, 7 and 14 days, cell viability was assessed utilizing the WST-1 cell proliferation reagent according to our previously published protocol [34 (link)] using a microplate reader (Model 3550, Bio-Rad Laboratories, Japan) at 450 nm with the reference wavelength of 650 nm. Gel discs without cells were used as negative controls.