Culturing Hela Cells

Watch the Hela cell culture protocol from thawing to plating out.



Medium: 90% of MEM or DMEM (with Earle's salts) with 10% FCS + 2 mM L-glutamine + non-essential amino acids. Can also use RPMI-1640 and 5-10% FBS.

Subculture: split confluent culture 1:4 or 1:6 every 3-5 days depending on confluency using trypsin/EDTA. Rate of doubling time is 24 to 48 hours. Seed at 1-2 x 106 cells/80 cm2. Incubate at 37 °C with 5% CO2

Storage: frozen with 70% medium, 20% FCS with 10% DMSO at about 1 x 106cells/ampoule

Hela cell culture protocol

Hela cells can be grown to the appropriate density usually 70% in a humidified chamber at 37°C, 5% CO2. HeLa cells in many labs are cultured in 75 cm2 flasks (or medium flasks). HeLa cells are grown in Dulbecco's Modified Eagle Medium with10% fetal bovine serum (FCS) and 1% MEM non-essential amino acids with penicillin-streptomycin at 1% is added to the culture media.

They can also be maintained in RPMI1640 containing penicillin-streptomycin, nonessential amino acids, sodium pyruvate, L-glutamine and 10% fetal calf serum and incubated in a humidified atmosphere with 5% CO2 at 37°C.