Freezing HeK'n cells
Posted 28 January 2005 - 04:40 AM
How can I freezzing HeK'n cells? I don't have special medium and I can't find no additional informations . I'll be grateful for any ideas.
Posted 31 January 2005 - 06:45 AM
Trypsinize the cells
Resuspend them in 'freezing buffer' at about 1 million cell/ml.
Freeze them slowly down to -80°C (put the vials between two styrofoam racks taped tight... this way they don't freeze too quickly).
The day after, put them in liquid nitrogen.
Freezing buffer : DMEM (or any medium I guess) + 20% FBS + 10% sterile DMSO. NO ANTIBIOTICS.
Posted 31 January 2005 - 08:36 AM
for a 15cm diameter plate :
i trypsinise cells, centrifuge them and add 2ml of a [SERUM / 10%DMSO]solution
Then i but cells in cryotubes. Cryotubes are putted in polystyrene racks taped tight and put overnight at -80°C. The second day i freeze them in nitrogen.
i prepare 9ml SERUM + 1ml sterile DMSO and store it for max 2weeks at 4°C
2ml are used for a 15cm diameter plate where cells are 80% confluent.
No less efficiency if the plate is full.
experiments done on hela cells, hek 293 and hek 293pack cells