Quantitative proteomics cell bilayer (qPCB) method for studying cell-cell communication

Cellular communication is a fundamental process in biology. We developed an in vitro procedure for quantitatively analyzing proteome-wide changes triggered by interaction of different cell types. Using an adipocyte-macrophage bilayer co-culture model we detected thousands of proteins and deciphered regulatory pathways involved in low-grade inflammation leading to insulin resistance. The method can be applied for multiple cell-cell combinations to gain understanding in cellular interaction. 2 cell lines (3T3-L1 adipocytes [A] and RAW 264.7 macrophages [R]), cultivated both as monolayer (condition 1 [O]) and together in a bilayer experiment (condition 2 [B]); 2-3 biological replicates per treatment (10 samples in total)