Microencapsulated 3D co-cultures recapitulate phenotypic features of B lymphomas

Non-Hodgkin B-cell lymphomas (B-NHL) mainly develop within lymph nodes (LN) as densely packed aggregates of tumor cells and their surrounding microenvironment, creating a tumor niche specific to each lymphoma subtypes. In vitro preclinical models mimicking biomechanical forces, cellular microenvironment, and 3D organization of B-cell lymphomas remain scarce, while all these parameters constitute key determinants of lymphomagenesis and drug resistance. Using a microfluidic method based on cell encapsulation inside permeable, elastic, and hollow alginate microspheres, we developed a new tunable 3D-model incorporating lymphoma B cells, extracellular matrix and/or stromal cells.Using RNA-sequencing, we reported that, in contrast to classical 2D cocultures, our 3D lymphoma model initiated a coevolution of these two cell types, recapitulating the physiopathology of B lymphoma tissue. DOHH2 and HLY1 cell lines were cultured in 2D or 3D conditions (alginate spheroids) with or without tonsil stromal cells (TSC) before cell sorting of CD20pos B cells and RNAseq