Human induced pluripotent stem cell culture

Coat 6 well plates with Geltrex LDEV-Free Reduced Growth Factor Basement Membrane Matrix (Thermo Fisher Scientific), 1:100 dilution for 1 hour before cell passaging.

When hiPSCs are 80-90% confluent, they are passaged:

Old media is aspirated and replaced with 1ml of cell-culture PBS (Gibco, Thermo Fisher Scientific)

PBS is aspirated and replaced with 0.5 mM EDTA (Thermo Fisher Scientific) made up in PBS.

hiPSC are incubated in EDTA for 0h 5m 0s at 37°C .

After 5 minutes, the EDTA is aspirated and cells are dissociated with media (E8 or mTesR 1).

hiPSCs are then placed in new 6 well plates at the desired dilution.

All hiPSCs are fed daily with fresh media. All cells were maintained at 37°C and 5% carbon dioxide.