Hi everyone!

I want to isolate the PBMCs from AML patients and transduce the cells with my lentivirus to knockdown a protein and use for further experiments. I am unable to find a single widely used and standardized protocol for short term liquid culture of AML blast cells, since in every paper people use different combination of cytokines. Another option I see is to use the StemSpan serum-free media with the CC100 cytokine cocktail. This could also work for me but it am trying to look for a more affordable option since this product CC100 is quite expensive. I do not wish to expand the cells and I work with the whole PBMC population (I am interested in the entire blast cell population) of different subtypes of AML patients, so if anyone is familiar with a protocol that is not specific for a particular AML subtype and is already using it successfully, I would be glad if you could share it!

I have used this media once that I read in a paper: RPMI with 10% FCS, 1% Pen-Strep supplemented with cytokines hIL3 (20ng/ml), hSCF (20ng/ml) and hGM-CSF (100ng/ml). The cells were more or less fine, but it was not a widely cited paper and I did not find this combination used by many people so I'm afraid it might not be the best option.

Would be glad to have your insights in this regard! Thank you!