I would like to make a kinetic analysis after drug release with short time points. We use RAJI cells that are not adherent, and we need a lot of cells (10xe8) for each time point (2', 5', 10', 15', ...). Centrifugation for the first wash is too long and we overpass the first time point. We thought on using filtration apparatus (the on we use for the serum). Does anyon have tried this yet ? Or have another idea to fastly concentrate cells ?
Thanks for you comments and answers
Please visit this web site
http://www.freepatentsonline.com/7291450.pdf
Regards
I think vacuum filtration sounds feasible, although the speed of it will depend on how much volume you have to process. Keeping the initial volume to a minimum will be important.
Also, you have to think about how you will collect the cells off of the filter. If you use the type of vacuum device in which the filter is separate from reservoir and held in place by a clamp, collecting the cells would be easier than if you use a filter unit with built-in membrane.
A more complicated method that has been used to obtain good time resolution is silicone oil centrifugation, in which the cells are spun through a layer of oil to separate them rapidly from the medium. This is normally done in quite small volumes, however, so might not be suitable for your experiment.
Thanks for your answers,
Karima, I need to separate my cells fastly to be treated in few minutes so I don't see how the patent could help but thanks;
Adam, yes I will probably treat huge volumes of cells because after the different time points I will have to make ChIP. I will try vaccum filtration.
Best
Perhaps you can pre-concentrate your cell suspension before beginning the treatment, in order to make the final filtration step faster.
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