The 92.1 primary human uveal melanoma cell line [14], kindly provided by Dr. Antonia Saornil from the Instituto Universitario de Oftalmobiología Aplicada (IOBA), University of Valladolid, was used. This selection was based on previous studies performed in our laboratory where this cell line demonstrated high proliferative and invasive potential

in vitro [15]. The cells were maintained at 37°C in a humidified 5% CO2-enriched atmosphere (Thermo Forma Series II Water Jacketed CO2 Incubator, Fisher Scientific Limited, Ontario, Canada). The cells were cultured in RPMI-1640 medium (Invitrogen, Burlington, Ontario, Canada), supplemented with 5% heat inactivated fetal bovine serum (FBS; Invitrogen), 1% fungizone (Invitrogen), and 1% penicillin-streptomycin (Invitrogen). One million cells (cellular viability greater than 99%) suspended in 0.1 ml of RPMI-1640 media were injected into the suprachoroidal space of the right eye of each rabbit according Selleck TSA HDAC to a previously described technique [13]. Ketamine (35 mg/kg; Vetalar, Vetrepharm Canada Inc., Belleville, Ontario, Canada)

and xylazine (5 mg/kg; Anased, Novopharm Limited, Toronto, Ontario, Canada) were used as anesthetics NSC23766 during the surgical procedure. Blue Light Exposure The 20 rabbits used in this experiment were randomly divided into two separate groups of 10 rabbits each. The experimental group was exposed to blue light 8 hours per day for the duration of the 8-week experiment. The animals were group-housed in a large pen into which the blue light-emitting apparatus was placed. Emricasan order heptaminol The

apparatus consisted of a large metal cage in which twenty-four 6600 k bulbs were suspended, each covered by a sheet of co-extruded polycarbonate film (Rosco, Color Filter #74 Night Blue) that allowed light only in the blue portion of the spectrum to pass through. This apparatus was placed in the middle of the pen, with suspended bulbs reaching to approximately 6″” from the ground to achieve maximal light exposure at eye level. Additionally, the pen was lined with 3′ high reflective aluminum to ensure adequate blue light exposure in all areas of the pen. As a rabbit’s gaze is typically 10 to 15 degrees below the horizontal plane, 3′ high reflective aluminum was adequate to ensure continuous blue light exposure in the direction of gaze. All lights were connected to a timer that turned on at 11 am and turned off at 7 pm daily. Protective goggles were provided to all personnel entering the housing area during the period of blue light exposure. The control group was in the adjacent pen, which was covered by a polycarbonate film (Rosco, Color Filter #15 Deep Straw) that ensured proper blockage of any light within the blue portion of the visible spectrum (500-444 nm, CIE International Diagram for blue light ranges) from entering the control pen. Fundoscopy Indirect ophthalmoscopy of dilated pupils using Tropicamide (Alcon Canada Inc., Mississauga, Canada; Mydriacyl, Alcon Canada Inc.