Cells - IGR3
Morphology: undifferentiated cells appear epithelial with short extensions. FluidRenewal: twice weekly
SubCulturing: Remove medium, rinse cell sheet with 0.25% trypsin, and add 1 to 2ml of fresh trypsin. Let the flask sit at room temperature until the cells detach. Add fresh medium, aspirate and dispense into new flasks.
SplitRatio: A ratio of 1:3 to 1:6 is recommended
Growth Properties: monolayer
Succinyl concanavalin-A Induced Differentiation of RGC-5: Cells cultured in the absence of serum for 24 hours. Then cultured in growth medium supplemented with succinyl concanavalin A (sConA; 50 µg/mL; Sigma) for 7 days. (stock 500x sConA 25mg/ml in DMEM -20C) Incubation of the cells with sConA induced differentiation and rendered the cells responsive to glutamate toxicity.