ARPE-19 Cell Culture and Characterization

ARPE-19 cells were obtained from the American Type Culture Collection (ATCC) and used for all experiments. Culture supplies were acquired from GE Healthcare for DMEM/F-12 media and Dulbecco’s phosphate-buffered saline (DPBS), ThermoFisher Scientific and Corning for culture flasks, VWR Seradigm for fetal bovine serum (FBS), and Corning for 24-well Transwell polyethylene terephthalate (PET) support membranes with 0.4 μm pores. Kits and reagents were obtained from ThermoFisher Scientific for the PicoGreen dsDNA assay kit, conjugated ZO-1 and F-actin antibodies, and NucBlue stain and from Sigma-Aldrich for the fluorescein isothiocyanate–dextran (FITC–dextran) molecules. Synthetic spider silk proteins were obtained through a previously described purification method of milk generated by transgenic goats that produce the dragline spidroins rMaSp1 and rMaSp2.^{67 (link)}