Herreros-Pomares Alejandro, Zhou Xuan, Calabuig-Fariñas Silvia, Lee Se-Jun, Torres Susana, Esworthy Timothy, Hann Sung Yun, Jantus-Lewintre Eloísa, Camps Carlos, Zhang Lijie Grace
Mixed Unit TRIAL, Fundación Investigacíón Hospital General Universitario de Valencia & Centro de Investigación Príncipe Felipe, Valencia, Spain; CIBERONC, Valencia, Spain.
Department of Mechanical and Aerospace Engineering, The George Washington University, Washington, DC, United States.
Mater Sci Eng C Mater Biol Appl. 2021 Mar;122:111914. doi: 10.1016/j.msec.2021.111914. Epub 2021 Jan 29.
Two-dimensional (2D) in vitro cell cultures and laboratory animals have been used traditionally as the gold-standard preclinical cancer model systems. However, for cancer stem cell (CSC) studies, they exhibit notable limitations on simulating native environment, which depreciate their translatability for clinical development purposes. In this study, different three-dimensional (3D) printing platforms were used to establish novel 3D cell cultures enriched in CSCs from non-small cell lung cancer (NSCLC) patients and cell lines. Rigid scaffolds with an elevated compressive modulus and uniform pores and channels were produced using different filaments. Hydrogel-based scaffolds were printed with a more irregular distribution of pores and a lower compressive modulus. As a 3D model of reference, suspension spheroid cultures were established. Therein, cancer cell lines exhibited enhanced proliferation profiles on rigid scaffolds compared to the same cells grown on either hydrogel scaffolds or tumor spheres. Meanwhile, primary cancer cells grew considerably better on hydrogel scaffolds or in tumor sphere culture, compared to cells grown on rigid scaffolds. Gene expression analysis confirmed that tumor spheres and cells seeded on hydrogel scaffolds significantly overexpress most of stemness and invasion promoters tested compared to control cells grown in 2D culture. A different phenomenon was observed within cells growing on the rigid scaffolds, where fewer significant variations in gene expression were detected. Our findings provide strong evidence for the advantageous usage of 3D printed models, especially those which use GelMA-PEGDA hydrogels as the primary scaffold material, for studying lung CSCs. The results demonstrated that the 3D printed scaffolds were better to mimic tumor complexity and regulate cancer cell behavior than in vivo 2D culture models.
二维（2D）体外细胞培养和实验动物传统上一直被用作临床前癌症模型系统的金标准。然而，对于癌症干细胞（CSC）研究，它们在模拟天然环境方面存在显著局限性，这降低了它们用于临床开发目的的可转化性。在本研究中，使用不同的三维（3D）打印平台建立了富含非小细胞肺癌（NSCLC）患者和细胞系CSC的新型3D细胞培养物。使用不同的细丝制作了具有提高的压缩模量和均匀孔隙及通道的刚性支架。基于水凝胶的支架打印出的孔隙分布更不规则且压缩模量更低。作为3D参考模型，建立了悬浮球体培养物。其中，与在水凝胶支架或肿瘤球体上生长的相同细胞相比，癌细胞系在刚性支架上表现出增强的增殖特征。同时，与在刚性支架上生长的细胞相比，原代癌细胞在水凝胶支架上或肿瘤球体培养中生长得要好得多。基因表达分析证实，与在二维培养中生长的对照细胞相比，肿瘤球体和接种在水凝胶支架上的细胞显著过表达大多数测试的干性和侵袭促进因子。在刚性支架上生长的细胞中观察到了不同的现象，在那里检测到的基因表达显著变化较少。我们的研究结果为3D打印模型，特别是那些使用GelMA-PEGDA水凝胶作为主要支架材料的模型，在研究肺CSC方面的有利应用提供了有力证据。结果表明，3D打印支架比体内二维培养模型更能模拟肿瘤复杂性并调节癌细胞行为。
