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Adding drug to cultured cells


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#1 Sunnyside

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Posted 20 April 2010 - 01:45 PM

I need to drug my cells (HEK293). I will be trying different exposure times ranging from 2 hours-6 hours.
Do I simply add the drugs to the regular media, or do I need to do something fancy like leaving out the fbs?
None of the protocols I found say - they all just indicate "cells were exposed to x concentration of drug for x period of time".
Any help/advice appreciated.
Thanks

#2 than4

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Posted 20 April 2010 - 04:06 PM

You can do it a number of ways depending on ease for you. From the wording you have indicated, I would assume that previously the drugs were added to cells in full serum media.

You can either directly add the drug to your cells in their existing media or make up the correct concentration of drug in enough media to cover your cells and replace the old media with new media containing drug. This can be useful if you are using small wells or need to use really small amounts of drug.

I would treat your cells at different time points and collect all the cells at once, rather than the other way around.

Edited by than4, 20 April 2010 - 04:07 PM.


#3 bob1

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Posted 20 April 2010 - 04:10 PM

It depends on the drug you are using and what you are looking at...

Some drugs need to be made up in reagents other than medium/water such as DMSO or DMF or even methanol - for these you need to make the drug up at a concentration such that you can add it at its effective level without adding too much of the diluting compound.

If you are looking at something that involves the cell cycle, it may be useful to synchronise the cells by serum starvation, but you would normally add the serum back into the medium for the duration of the experiment, so that the cells are growing under "normal" conditions.

#4 Inmost sun

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Posted 22 April 2010 - 08:54 AM

I need to drug my cells (HEK293). I will be trying different exposure times ranging from 2 hours-6 hours.
Do I simply add the drugs to the regular media, or do I need to do something fancy like leaving out the fbs?
None of the protocols I found say - they all just indicate "cells were exposed to x concentration of drug for x period of time".
Any help/advice appreciated.
Thanks


it is a good idea to compare with and w/o FCS; FCS may immobilze your drug if it is amphi- or lipophilic; but w/o you reduce triggering survival signalling pathways...




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