I need to sort 1-2 million dendritic cells for a proteomic group. We would like to sort these DC into serum / protein free solution, but the DC all die very quickly and we do not recover any cell pellet after sorting even 1 Mio DC. I get a pellet when sorting into medium containing 10% serum. But the proteomic people don´t like FBS as it is interfering with down stream analysis (get a lot of FBS proteins in the mass spec). I´ve sorted into medium with serum and then washed serum/protein free. Again all DC`s are efficiently killed and nothing recovered after centrifugation. At the moment I´ve sorted using the 70 mikroM nozzle and I will test the 100 mikroM. But that will increase the volume of my sort, which is already huge (1.5 ml fluid/ 750 000 cells). Any better solutions? Thanks for all thoughts and hints.
Make sure that no Na azide in your buffers and use PBS with 0.5% BSA (prevent cell adsorption) you may wash once after that in PBS.
Either you use Serum Free medium
https://www.thermofisher.com/uk/en/home/life-science/cell-culture/mammalian-cell-culture/serum-free-media.html
look at my mass spect papers on my page I already did proteomics on DCs without serum and it works really well.
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