A novel 3D culture model recapitulates primary FL B-cell features and promotes their survival

Abstract

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 (ECM), and/or tonsil stromal cells (TSC). We revealed that under 3D confinement lymphoma B cells were able to form cohesive spheroids resulting from overexpression of ECM components. Moreover, lymphoma B cells and TSC dynamically formed self-organized 3D spheroids favoring spheroid growth. 3D culture induced resistance to classical chemotherapeutic agent doxorubicin, but not to BCL2 inhibitor ABT-199, identifying this approach as a relevant in vitro model to assess the activity of therapeutic agents in B-NHL. RNAseq analysis highlighted the synergy of 3D, ECM, and TSC in upregulating similar pathways in malignant B cells in vitro than those overexpressed in primary lymphoma cells in situ. Finally, our 3D model including ECM and TSC allowed long-term in vitro survival of primary follicular lymphoma B cells. In conclusion, we propose a new high throughput 3D model mimicking lymphoma tumor niche and making it possible to study the dynamic relationship between lymphoma B cells and their microenvironment and to screen new anti-cancer drugs.

Application of 3D Hepatic Plate-Like Liver Model for Statin-Induced Hepatotoxicity Evaluation

“…hy926 cell line induced an anisotropic intercellular mechanical tension which is crucial in inducing hepatocyte polarization. It has been reported that ECM can be contained in Na-Alg hydrogel and influence cell polarity ( Lamaison et al, 2021 ; Doméjean et al, 2016 ), which is consistent with our findings. In the 3D-P co-culture system, FN distributed on both sides of hepatocytes and might induce polar intercellular mechanical tension and further guide the formation of bile canaliculus by mimicking the ECM in the space of Disse, which will not occur in most of the nonpolar 3D co-culture systems.…”

Development and Characterization of 3D Hybrid Spheroids for the Investigation of the Crosstalk Between B-Cell Non-Hodgkin Lymphomas and Mesenchymal Stromal Cells

“… 38 Notably, in our study, no protective effect of MSCs in 3D DLBCL culture was observed, which is in line with the recent report of Lamaison et al performed on DLBCL and follicular lymphoma (FL) spheroids. 69 Importantly, hybrid models described here share other characteristics with DLBCL spheroids developed by Lamaison and co-authors. First, we observed a similar survival decrease during the second week of culture; however, the rate of cell death of our model was significantly lower.…”

In Vitro and In Vivo Models of CLL–T Cell Interactions: Implications for Drug Testing

“…A dynamic system might also mimic CLL interactions in bone marrow by introducing scaffolds coated with bone-marrow stromal cells (HS5 cell line) [126]. Cultures incorporating T cells or even complex lymphoid structures have not yet been reported for CLL, but 3D culture in a microfluidic system enables the in vitro survival of follicular lymphoma primary cells co-encapsulated with tonsil stromal cells [127].…”