Culture conditions for cell treatments - (Sep/10/2009 )
I'm going to do some experiments on a cell model treated with a specific compound. The problem is that such compound is contained in standard cell culture medium, so you think I simply can use a higher concentration to investigate the effects?
Also: I know cells are usually serum starved for 24 hr before treatments to synchronize cell cycles. Is that a rule or I can bypass this step?
And what regards the serum content of medium used to dissolve the compound and do treatments? I have to completely eliminate serum also during cell treatment?
I hope someone can help me!!!
Thanks in advance for your replies!
It depends... is the factor critical for the cells to survive? If so, you may want to consider going over to one of the serum free mediums (Invitrogen produce a range), or getting a custom made medium (expensive).
Serum starvation can be used to synchronize the cells, it is the easiest and cheapest way to do so. There are other methods, but they involve drug treating the cells - which then has other off target effects.
I do a similar expt.....where i increse one of the compounds already present in the media...just do these treatment in absence of serum...first try out in the range of conc starting from the conc presnt in the media and with the difference of 0.5 or 1 unit ...try diff conc at one go in 6-well plate. try these in media with LG and serum free.u can use antibiotic if reqiured.i use it.
Use varying concentrations of extract and then treat with a reagent called WST-1. Use the data to generate a curve of cytotoxicity and use the highest concentration which remains uncytotoxic.
Thank you so much for your replies!
I'll not use serum during treatments as you proposed and try to define the right concentration by a cell viability test.
All the best!