Hi, I think you can use the protocols which are based on blood serum or blood agar media as all the essential nutrients will be available other than that it wont stick to the plate too.

Hi Davide,

I routinely make bone-marrow derived macrophages with 20 ng/mL GM-CSF and 100 ng/mL IL-4 in RPMI 1640 buffered with 10 mM HEPES. I allow these to mature for 6 days, changing the media once at day three and replacing with fresh cytokines. This produces immature dendritic cells, which I then stimulate with 1 ng/mL LPS for 24 hours. 

I have never personally made DC's from splenocytes, but from the protocols I have seen, the cytokine concentrations used are similar to the ones I use reliably for BMDC's.

Others in my lab have pulsed DC's with tumor cell conditioned media supernatants for 3 days. Hope this helps!

Hi Valerie,

thanks for your answer, it is indeed really helpful.

I just have one more "technical" question. When you change the media, do you harvest and centrifuge the supernatant with cells, and then replace it? Or what do you do in order to not "touch" too much the DCs?

Thanks again!