Comparison of 2D and 3D cell culture models for cell growth, gene expression and drug resistance
Comparison of 2D and 3D cell culture models for cell growth, gene expression and drug resistance
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DOI：
10.1016/j.msec.2019.110264
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发表时间：
2020-02-01
影响因子：
7.9
通讯作者：
Bonorino, Cristina
中科院分区：
文献类型：
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作者：
Fontoura, Julia C.;Viezzer, Christian;Bonorino, Cristina
体外药物筛选被广泛用于新药开发，因为它们构成了选择具有更大治疗潜力的化合物的具有成本效益的方法。它们也是体内测试的有吸引力的替代方案。然而，大多数这些测定是在二维培养模型中进行的，其中细胞在聚苯乙烯或玻璃平面上生长。为了开发更接近生理条件的体外模型，已经开发了三维模型。在这里，我们介绍了两种新型的全合成支架使用聚合物聚羟基丁酸酯（PHB）：溶剂浇铸颗粒浸出（SCPL）膜;和电纺膜，用于B16 F10小鼠黑色素瘤细胞和4 T1小鼠乳腺癌细胞的3D培养。使用常规组织培养板中的2D细胞培养系统和经典3D模型（其中细胞在源自Engelbreth-Holm Swarm（EHS）肿瘤的市售凝胶上生长）与合成支架进行比较。还从在同基因小鼠中作为移植物生长的体内肿瘤收集细胞。形态学，细胞活力，化疗反应和基因表达分析用于比较所有系统。在电纺膜模型中，细胞生长在纳米级纤维上，并且在SCPL膜中，其为细胞生长提供泡沫状结构，孔径变化。在所有3D模型上生长的细胞能够形成聚集体和球状体，与2D系统相比，允许增加细胞-细胞接触。细胞形态在3D系统和从体内肿瘤收集的细胞之间也更相似。在3D模型中生长的细胞显示出对达卡巴嗪和顺铂的抗性增加。基因表达分析也揭示了所有3D平台之间的相似性。这两种合成系统与经典EHS凝胶模型之间的相似性突出了它们作为药物筛选中具有成本效益的替代品的潜在应用，其中全合成模型可以代表迈向更高重现性的一步。我们得出结论，聚羟基丁酸合成膜提供了一个有价值的替代三维文化。
In vitro drug screening is widely used in the development of new drugs, because they constitute a cost-effective approach to select compounds with more potential for therapy. They are also an attractive alternative to in vivo testing. However, most of these assays are done in two-dimensional culture models, where cells are grown on a polystyrene or glass flat surface. In order to develop in vitro models that would more closely resemble physiological conditions, three-dimensional models have been developed. Here, we introduce two novel fully synthetic scaffolds produced using the polymer polyhydroxybutyrate (PHB): a Solvent-Casting Particle-Leaching (SCPL) membrane; and an electrospun membrane, to be used for 3D cultures of B16 F10 murine melanoma cells and 4T1 murine breast cancer cells. A 2D cell culture system in regular tissue culture plates and a classical 3D model where cells are grown on a commercially available gel derived from Engelbreth-Holm Swarm (EHS) tumor were used for comparison with the synthetic scaffolds. Cells were also collected from in vivo tumors grown as grafts in syngeneic mice. Morphology, cell viability, response to chemotherapy and gene expression analysis were used to compare all systems. In the electrospun membrane model, cells were grown on nanometer-scale fibers and in the SCPL membrane, which provides a foam-like structure for cell growth, pore sizes varied. Cells grown on all 3D models were able to form aggregates and spheroids, allowing for increased cell-cell contact when compared with the 2D system. Cell morphology was also more similar between 3D systems and cells collected from the in vivo tumors. Cells grown in 3D models showed an increase in resistance to dacarbazine, and cisplatin. Gene expression analysis also revealed similarities among all 3D platforms. The similarities between the two synthetic systems to the classic EHS gel model highlight their potential application as cost effective substitutes in drug screening, in which fully synthetic models could represent a step towards higher reproducibility. We conclude PHB synthetic membranes offer a valuable alternative for 3D cultures.