Culturing protocol:

- Use Eagle’s Minimum Essential Medium (DMEM and RPMI1640 also works well) supplemented with 10% FBS. Discard culture medium every 2-3 days

- To passage cells, briefly rinse cell monolayer with 1xPBS twice and add pre-warmed (37°C) 0.05% Trypsin-EDTA solution for 5 – 7 minutes

- Once cell layer is dispersed (5-7 min at 37°C) deactivate Trypsin by adding equal volume of complete growth medium with 10% FBS

- To avoid clumping don’t agitate the cells by shaking the flask while waiting for the cells to detach

- Split cells 1:4 every 3 days (recommended), or 1:8 every 6 days.

- Cultures should be incubated at 37°C in humidified atmoshere with 5% CO2

You can use the pdf file attached for more details.

thanks, Rafael but my question is cells is confluent ready to treat after 24 hr I have to add the drug in that can I use FBS & Streptomycin in media or only DMEM pl suggest me

If you wish to have useful advice, clearly state the purpose of your experiment. Do not expect others to be mind-readers.