Bioinstructive microparticles for self-assembly of mesenchymal stem Cell-3D tumor spheroids
Bioinstructive microparticles for self-assembly of mesenchymal stem Cell-3D tumor spheroids
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DOI：
10.1016/j.biomaterials.2018.09.007
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发表时间：
2018-12-01
期刊：
影响因子：
14
通讯作者：
Mano, J. F.
中科院分区：
文献类型：
--
作者：
Ferreira, L. P.;Gaspar, V. M.;Mano, J. F.
三维多细胞肿瘤球体（3D- mcts）在体外密切模拟复杂的肺肿瘤微环境（TME），是非常理想的筛选创新抗癌治疗方法。尽管在模拟肺TME方面取得了重大进展，但很少有模型将骨髓肿瘤浸润间充质干细胞（hBM-MSCs）与含有模拟ECM成分的异型3D肿瘤球体模型结合起来。在此，我们设计了混合3D-MCTS，首次结合了A549：成纤维细胞：异型三培养中的hBM-MSCs，以及生物指导性透明质酸微粒，作为肿瘤ecm模拟物和细胞锚定热点。结果表明，三维微球为细胞自组装成致密的三维微组织提供了适当的支持，并促进了CD44表达的增加，模拟了天然ecm透明质酸的存在。3D-MCTS的大小和球状形态可复制，三培养模型呈现典型的实体瘤坏死核心。间充质干细胞跟踪表明，hBM-MSCs在3D微肿瘤团块中迁移到不同区域，表现出与癌细胞和间质成纤维细胞的动态相互作用，与人类肿瘤相似。重要的是，阿霉素在三维三培养模型和hBM-MSCs:A549的双重培养中显示了hBM-MSCs对细胞毒性反应的影响，比例为10:1。这些发现证明了将hBM-MSCs与癌症间质成纤维细胞联合用于3D体外肿瘤模型的相关性，以及测试不同细胞间比以模拟肿瘤异质性的重要性。此外，具有生物指导性的透明质酸微粒也能有效地作为细胞凝聚支架，并显示出未来在体外3D模型中包含不同ECM成分的潜力。
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.