abstract
3D multicellular tumor spheroids (3D-MCTS) that closely mimic in vitro the complex lung tumor micro environment (TME) are highly desirable for screening innovative anti-cancer therapeutics. Despite significant improvements in mimicking lung TME, few models have combined tumor-infiltrating mesenchymal stem cells from bone marrow (hBM-MSCs) with heterotypic 3D tumor spheroid models containing ECM mimetic components. Herein, we engineered hybrid 3D-MCTS that combine, for the first time, A549:fibroblasts:hBM-MSCs in heterotypic tri-culture, with bioinstructive hyaluronan microparticles that act as tumor-ECM mimetics and as cell-anchoring hotspots. The obtained results indicated that 3D microspheres provided proper support for cells to self-assemble into compact 3D microtissues and promoted an increase in CD44 expression, emulating the presence of native-ECM hyaluronan. 3D-MCTS size and sphere-like morphology was reproducible and tri-culture models presented the characteristic solid tumors necrotic core. Mesenchymal stem cells tracking demonstrated that hBM-MSCs migrate to different regions in 3D microtumors mass exhibiting dynamic interactions with cancer cells and stromal fibroblasts, alike in human tumors. Importantly, doxorubicin administration revealed hBM-MSCs effect on cytotoxic responses in 3D tri-culture models and in dual cultures of hBM-MSCs:A549 at 10:1 ratio. Such findings evidence the relevance of including hBM-MSCs in combination with cancer-stromal fibroblasts in 3D in vitro tumor models and the importance to test different cell-to-cell ratios to mimic tumor heterogeneity. In addition, bioinstructive hyaluronan-microparticles were also effective as cell-agglomerating scaffolds and showed potential to be used as an enabling technology for including different ECM components in 3D in vitro models in the future.
keywords
LUNG-CANCER CELLS; DOUBLE-EDGED-SWORD; HYALURONIC-ACID; MULTIDRUG-RESISTANCE; 3D; CULTURE; MICROENVIRONMENT; FIBROBLASTS; EXPRESSION; GROWTH
subject category
Engineering; Materials Science
authors
Ferreira, LP; Gaspar, VM; Mano, JF
our authors
João Mano
Vice Director
Luís Pedro Correia Pinto Ferreira
PhD Student
Vítor Gaspar
Assistant Professoracknowledgements
The authors would like to acknowledge the support of the European Research Council grant agreement ERC-2014-ADG-669858 for project "ATLAS". The authors also acknowledge the financial support by the Portuguese Foundation for Science and Technology (FCT) through a Post-doctoral grant (SFRH/BPD/119983/2016, Vitor Gaspar). This work was also developed within the scope of the project CICECO - Aveiro Institute of Materials, POCI-01-0145-FEDER-007679 (FCT Ref. UID/CTM/50011/2013), financed by national funds through the FCT/MEC and when appropriate co-financed by FEDER under the PT2020 Partnership Agreement.