Cell lines are provided either as frozen cultures (in plastic cryo-ampoules sent on dry ice) or as active (growing) cultures in plastic flasks or tubes. Frozen cells should not be stored at -80 ̊C, but in liquid nitrogen (-196 ̊C). Please take precautions when handling dry ice or liquid nitrogen. Direct skin contact may cause severe injuries. Plastic or glass ware may burst or even explode upon thawing.
For the first few days we recommend use a higher concentrations of fetal bovine serum (FBS) than usual. Routine supplementation with antibiotics can mask contaminations and should be avoided. Unless stated otherwise, incubate and propagate the cells at 37 ̊C in an humidified atmosphere containing 5% CO2. Do not forget to freeze reserve ampoules as soon as possible as backup and for replication. For further information or specifics please contact our customer support.
Thaw the ampoule in a waterbath (25-37 ̊C). Transfer the cells to a 10-15 ml centrifuge tube. Add 6-10 ml prewarmed medium to the cells, mix gently (some scientists recommend dropwise addition of medium) and remove a small sample to determine cell number and viability. Centrifuge at 200 x g for 5 min. Decant supernatant and adjust the cell suspension to the recommended concentration in the final culture medium (as outlined on the cell line data sheet). Seed out in a flask or plate of suitable size according to the instructions given for the respective cell line. Culture in Petri dishes is not recommended. We recommend to start the culture with 20% FBS and once the cells show vigurous growth to decrease the FBS concentration to lower concentrations.
Active Culture - Monolayer
The culture vessel is usually completely filled with medium for transport. Carefully remove transport medium and add freshly prepared, prewarmed culture medium sufficient to cover the bottom of the flask. Incubate and subdivide the culture as outlined in the cell line information sheet. If the cells detach from the plastic surface in transit, remove the entire contents of the flask and centrifuge at 200 x g for 5 min. Discard the supernatant and resuspend cells in fresh medium (after removing an aliquot to determine cell number and viability). Seed the cell suspension in a flask of suitable size according to the instructions given for the respective cell line.
Active Culture - Suspension Culture
For shipment the flask or tube was completely filled with medium. Carefully remove transport medium with cells and centrifuge at 200 x g for 5 min. Resuspend the cells in a defined volume of medium and determine cell number and viability. Resuspend the cells in freshly prepared culture medium containing the required supplements and incubate as recommended in the cell line information sheet.