I want to grow Panc-1 cells together with PBMCs in an 'indirect' co-culture model, for anywhere up to 72 hours, with the eventual goal of assessing changes in cyto/chemokine expression and viability (of the Panc-1 cells) in response to drug treatment.

We are in the process of developing a protocol for this co-culture, and my initial plan was to use a transwell system with 0.4um pores. Panc-1 is ofc an adherent cell line. My understanding of PBMCs is that adherence triggers differentiation into macrophages, which for our purposes we want to try and avoid.

I'm trying to understand if I should seed the panc-1 cells in the upper compartment of the insert and use an ultra-low adherent well to culture the PBMCs below, or the reverse, and use a TC treated 24 well plate for the Panc-1 and place the PBMCs in the upper compartment? I have access to both polyester and polycarbonate membrane inserts.

Placing panc-1 in the upper compartment complicates things in terms of assessing viability (which we've thus far done using fluoresence techniques, e.g. SRB Assay), so isn't my first choice.