A53T aSYN-IRES-GFP Stable LUHMES Cell Line

Grow the cells in culture vessel pre-coated with 50 μg/ml poly-L-ornithine (PLO) (TM062) and 1 μg/ml fibronectin (EMD Millipore; Cat. FC010) in H2O for at least 3 hours at 37°C. Do not grow these cells in culture vessels with surface areas equal to or less than 12.5 cm2. These cells do not grow in 6-well

24-well

48-well

or 96-well plates.  Advanced DMEM/F12 + 1X N2 supplement (ThermoFisher Scientific) + 2mM L-glutamine (G275) + 40ng/ml Recombinant Human FGF2 (Z101455). Carbon dioxide (CO2): 5%

Temperature: 37.0°C. Note: Cells may be grown in the presence of 1 μg/ml Tetracycline. Change media every 2-3 days; do not let media change colour to orange-yellow. The cells will form round clumps instead of a monolayer when stressed. Avoid subculturing if the cells appear stressed. If leaving the cells over the weekend (or more than 2 days)

split cells at high ratio (1:4 to 1:5). Cells are sensitive to trypsin; Gentle Dissociation Solution (TM080) is recommended for subculture procedures. For information regarding cell differentiation

please refer to the Differentiation Protocol PDF under the Documents Tab.