The Effect of Increased Pressure on In Vitro Survival of Transfected Rat Retinal Ganglion Cells

Abstract

Abstract: : Purpose: To study the effect of varying pressure on the survival of retinal ganglion cells(RGC) at differential time intervals in a closed chamber pressure system. Methods: Rat retinal ganglion cells (RGC–5) were cultured using Delbecco's minimum essential medium at 37°C for 3 days. The cells were counted before and after the experiment. After a monolayer was established with 80% confluence, the culture was transferred into an incubation chamber (Solent Scientific) coupled with a Constar temperature videomicrography system. Cells were exposed to pressure of 30 and 75 mm Hg. Cells were monitored continuously by phase–contrast time lapse videomicrography for 2, 6 and 18 hours at 37°C.Cell survival was assessed with flow cytometry and annexin staining Results: The percentage of apoptotic cell–death stained with Annexin–V at differential pressure and time are depicted in Table 1. Rate of apoptosis linearly correlated with degree of pressure and duration of exposure (p<0.01) Conclusions: Our results suggest that pressurization induces apoptosis in a time dependent manner and degree of cell death is a function of higher pressure. This system is an ideal model to study the effects of various neuroprotective agents on the survival of ganglion cells under high pressure. . Table 1 

Keywords: apoptosis/cell death • ganglion cells