Moving S2 cells from one plate to another
Posted 15 June 2010 - 06:16 AM
Posted 25 June 2010 - 12:00 PM
Hello, I have seeded Drosophilia S2 cells from a main 75 cm2 flask into 6 well plates at a cell concentration of 5x10^6 cells/ml with 2ml/well (1x10^7 cells/well). I let the cells sit in the wells for 30 minutes before adding a drug to them. Is 30 minutes a sufficient amount of time for the cells to get acclimated to their new surroundings before adding a drug or should I have let them sit longer? The drug is a protein kinase B inhibitor and it doesn't hurt the cells too much.
are they suspension? if you are only changing the physical environment (wells, temperature from leaving incubator...) the day of the experiment, i would not wait much time at all after i changed wells... simply to get them back in the incubator faster. i suppose you could change the well then re incubate them for 30 minutes before adding the drug, but honestly if it were me i would not take my plates in and out of the incubator/hood and removing the plate cover often, just increases chances of infection. I would have all my media/plates prepared, remove from incubator and aliquot into the wells that have the conditions (drug or controls) and place back in incubator if that makes sense. just a noob here tho mayb others give u more advice with this bump