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ACCEPTED MANUSCRIPT (n=5); PD only (n=5); PD + 670nm (n=5). The animal received 670nm light prior to PD, according to previously described methods (Albarracin et al., 2011; Albarracin and Valter, 2012; Jager et al., 2008; Rutar et al., 2010). PD was induced by 2x36W COLDF2 fluorescent 2 tubes, as described previously (Fernando et al., 2016) delivering 1000lux (147uW/cm ) power density. Whole eyes and retinas were immediately collected after PD for histological analysis and RNA extraction. Five animals were used for each experimental group. 2.2 Maintenance of 661W photoreceptor-like cells Murine photoreceptor-derived 661W cells were kindly gifted by Dr. Muayyad R. Al- Ubaidi (Department of Cell Biology, University of Oklahoma Health Sciences Centre, Oklahoma City, OK, USA). Cells for experimental purposes were used within five passages of authentication, and validation of authenticity was performed using gene expression of green cone pigments and cone arrestin. Cells were further validated for species authenticity (CellBank, Sydney, Australia). Cells were cultured in growth medium (GM) containing Dulbecco’s Modified Eagle Medium (DMEM; Sigma-Aldrich, MO, USA) supplemented with 10% fetal bovine serum (FBS; Sigma-Aldrich), 6mM L-glutamine (Thermo Fisher Scientific, MA, USA) and antibiotic-antimycotic (100U/ml penicillin, 100μg/ml streptomycin and 0.25μg/ml Fungizone; Thermo Fisher Scientific), and incubated in dim conditions in a humidified atmosphere of 5% CO2 at 37°C in the dark. 2.3 Preparation of rat primary Müller cells Retinas were isolated from 36 SD rats aged P8-10 days following the modified methods described previously (Hicks and Courtois, 1990). Isolated retinas were immersed in DMEM supplemented with 6mM L-glutamine and antibiotic-antimycotic for 2 hours at 4°C. Retinas were dissociated into small fragments and then incubated with DMEM containing 0.1% trypsin, 70U/ml collagenase type IV, and antibiotic-antimycotic for 1 hour at 37°C. Cell 6 ACCEPTED MANUSCRIPTPDF Image | Photobiomodulation with 670 nm light ameliorates retinal
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