Simultaneous 2D and 3D cell culture array for multicellular geometry, drug discovery and tumor microenvironment reconstruction
Simultaneous 2D and 3D cell culture array for multicellular geometry, drug discovery and tumor microenvironment reconstruction
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用于多细胞几何、药物发现和肿瘤微环境重建的同时 2D 和 3D 细胞培养阵列
DOI：
10.1088/1758-5090/ac1ea8
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发表时间：
2021-10-01
期刊：
影响因子：
9
通讯作者：
Liu, Yiyao
中科院分区：
文献类型：
--
作者：
Li, Shun;Yang, Kaifu;Liu, Yiyao
细胞培养系统是生物医学研究中不可缺少的体外工具。虽然传统的二维（2D）细胞培养仍然用于大多数生物医学和生物学研究，但三维（3D）细胞培养技术吸引了越来越多的研究人员的关注，特别是在癌症和干细胞研究中。由于不同的空间结构，2D和3D培养物中的细胞表现出不同的生物化学和生物物理表型。因此，需要具有2D和3D细胞培养物的新平台来弥合2D和3D基于细胞的测定之间的差距。在这里，同时二维和三维细胞培养阵列系统构建的显微打印技术，其中癌细胞表现出杂合的几何结构与二维单层和三维球体。三维球体中生长的细胞具有更高的增殖能力和更强的细胞-细胞粘附。来自不同类型的癌细胞系的球体通过几何约束刺激的生物力学转导表现出不同的形态。癌细胞聚集体的Z投影图像用于分析3D多细胞结构特征。值得注意的是，通过使用支持向量机分类器，我们区分肿瘤细胞与正常细胞的准确性大于95%，根据多细胞球体的几何特征，在相差显微镜图像。多细胞球体阵列中的癌细胞比2D培养中生长的细胞表现出更高的抗癌药物顺铂的耐药性。最后，我们建立了一个由肿瘤球体阵列、成纤维细胞和光交联明胶甲基丙烯酰水凝胶组成的共培养系统，以模拟由实体瘤团块、周围基质细胞和细胞外基质组成的肿瘤微环境。总之，我们新开发的同时2D和3D细胞培养阵列在2D和3D细胞事件的综合评估、球体阵列的快速生产和基于多细胞几何学的肿瘤细胞检测方面具有巨大潜力。
Cell culture systems are indispensable in vitro tools for biomedical research. Although conventional two-dimensional (2D) cell cultures are still used for most biomedical and biological studies, the three-dimensional (3D) cell culture technology attracts increasing attention from researchers, especially in cancer and stem cell research. Due to the different spatial structures, cells in 2D and 3D cultures exhibit different biochemical and biophysical phenotypes. Therefore, a new platform with both 2D and 3D cell cultures is needed to bridge the gap between 2D and 3D cell-based assays. Here, a simultaneous 2D and 3D cell culture array system was constructed by microprinting technology, in which cancer cells exhibited heterozygous geometry structures with both 2D monolayers and 3D spheroids. Cells grown in 3D spheroids showed higher proliferation ability and stronger cell-cell adhesion. Spheroids derived from various types of cancer cell lines exhibited distinct morphologies through a geometrical confinement stimulated biomechanical transduction. Z-projected images of cancer cell aggregates were used to analyze 3D multicellular architecture features. Notably, by using a support vector machine classifier, we distinguished tumor cells from normal cells with an accuracy greater than 95%, according to the geometrical features of multicellular spheroids in phase contrast microscopy images. Cancer cells in multicellular spheroid arrays exhibited higher drug resistance of anticancer drug cisplatin than cells grown in 2D cultures. Finally, we developed a co-culture system composed of tumor spheroid arrays, fibroblast cells and photo-crosslinkable gelatin methacryloyl hydrogel to mimic tumor microenvironment which consisted of solid tumor massed, surrounding stromal cells and extracellular matrix. Together, our newly developed simultaneous 2D and 3D cell culture array has great potential in comprehensive evaluation of cellular events in both 2D and 3D, rapid production of spheroid arrays and multicellular geometry-based tumor cell detection.