All chemicals were purchased from Sigma–Aldrich (St. Louis, MO, USA) and used as received unless otherwise described. Curcumin (368.38g/mol; ≥94% Curcuminoid content) was obtained from Sigma. Propylene sulfide was purchased from Acros Organics (NJ, USA) and purified by distillation just before polymerization. 87–90% hydrolyzed poly(vinyl alcohol) (PVA) of average molecular weight 30,000–70,000 was prepared into a 1% w/v solution in deionized water. Transwell inserts with 0.4 μm pore polycarbonate membranes (Corning, Lowell, MA, USA) were used in 24 well plates for Curcumin release experiments. SIN-1 was purchased from Invitrogen (San Diego, CA, USA) as a package of 1 mg vials. Cell culture reagents, including fetal bovine serum (FBS), Dulbecco’s Modified Eagle Medium (DMEM), and penicillin-streptomycin (p-s) were supplied by Gibco Cell Culture (Carlsbad, CA, USA). 4-Cyano-4-(ethylsulfanyltiocarbonyl) sulfanylpentanoic acid (ECT) was synthesized following the previously reported procedure [40 (link)].
Transwell inserts with 0.4 μm pore polycarbonate membranes
Manufactured by Corning
Sourced in United States
Transwell inserts with 0.4 μm pore polycarbonate membranes are porous cell culture inserts designed for use in multiwell plates. The polycarbonate membrane has a pore size of 0.4 μm, enabling the study of cell migration, invasion, and permeability.
Automatically generated - may contain errors
2 protocols using transwell inserts with 0.4 μm pore polycarbonate membranes
1
Curcumin-loaded PVA Nanoparticles for Release
2
Measuring Transepithelial Electrical Resistance
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For the measurement of the transepithelial electrical resistance (TEER), 12 mm Transwell® inserts with 0.4 μm pore polycarbonate membranes (Corning) coated with collagen IV (20 μg/ml, Sigma-Aldrich) were used. OKG4 cells (2.7 × 104) in 500 μl in DermaLife K medium containing 60 μM Ca2+ were seeded to the filter, and the medium was changed every 1–3 days. When cells reached confluence (approx. after 7 days), they were cultured in a medium containing 1.4 mM Ca2+ to induce terminal cell differentiation [29 (link)] and the formation of tight junctions [30 (link)]. Subsequently, 50 μg/ml of the nanocarrier solution was applied to the cells. As a control, the TEER of cells that were kept in DermaLife K medium containing 60 μM Ca2+ was determined. The TEER values were measured using an Endohm-12 chamber (World Precision Instruments) and a volt-ohm-meter (Millipore). After subtracting the blank filter’s TEER, the value was multiplied by the filter area (1.12 cm2). Four independent experiments were performed in duplicate.
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