Grow HeLa in Eagle’s MEM modified with 10% FBS. Incubate at 37.0°C with 5% CO2. Renew growth medium 2-3 times a week
Subculture Protocol for HeLa:
- Rinse cells with 0.25% Trypsin/0.53mM EDTA
- Add 3 mL of Trypsin-EDTA to flask and watch for cell layer dispersal under an inverted microscope. this should occur within 5-15 minutes. Do not agitate cells during this type. Agitation encourages clustering. If cells are not detaching, place in incubator for 5 minutes to facilitate dispersal.
- Centrifuge cells and remove Trypsin-EDTA supernatant.
- Add 6-8 mL of growth medium and aspirate cells with pipette, gently.
- Add aliquots of cells suspension to culture vessels.
- Incubate at 37°C in 5% CO2.
A subcultivation ratio of 1:2 to 1:6 is recommended
Freeze cells in conditioned growth medium supplemented with 5% (v/v) DMSO and store in the liquid nitrogen vapor phase or at -135 °C.