HACAT Human Immortalized Epidermoid Cell Line

2-3 mL for T75 flask), 5) Place in a 37°C incubator for digestion for 1-2 minutes (difficult-to-digest cells can be properly prolonged for digestion time), 6) Observe the cell digestion under a microscope. If most of the cells become round and fall off, and add 3-4 ml of culture medium containing 10% FBS to terminate digestion. 7) Gently mix and aspirate, and perform subsequent passages at a ratio of 1:2~1:5 as per actual conditions., and rinse the cells 1-2 times with PBS without calcium and magnesium ions. 4) Add 0.25% (w/v) trypsin-0.53 mM EDTA to the culture flask (1-2 mL for T25 flask, centrifuge at 1000rpm for 3-5min, quickly take it back to the operating table, Recovery of Frozen Cells: For Suspension Cell passaging, subculture can be performed. For Adherent Cell passaging, tap the culture flask a few times