Cell Culture Information

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:

To subculture:

  1. Rinse cells with 0.25% Trypsin/0.53mM EDTA
  2. 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.
  3. Centrifuge cells and remove Trypsin-EDTA supernatant.
  4. Add 6-8 mL of growth medium and aspirate cells with pipette, gently.
  5. Add aliquots of cells suspension to culture vessels.
  6. Incubate at 37°C in 5% CO2.

A subcultivation ratio of 1:2 to 1:6 is recommended

To preserve:

Freeze cells in conditioned growth medium supplemented with 5% (v/v) DMSO and store in the liquid nitrogen vapor phase or at -135 °C.


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