Weiss Felix, Atlasy Nader, van Reijmersdal Vince, Stunnenberg Henk, Hulsbergen-Veelken Cornelia, Friedl Peter
Department of Cell Biology, Radboud University Medical Centre, P.O. Box 9101, 6525 GA Nijmegen, The Netherlands.
Department of Molecular Biology, Faculty of Science, Radboud University, 6525 AJ Nijmegen, The Netherlands.
In Vitro Model. 2022;1(6):463-471. doi: 10.1007/s44164-022-00040-x. Epub 2023 Mar 15.
3D in vitro culture models of cancer cells in extracellular matrix (ECM) have been developed to investigate drug targeting and resistance or, alternatively, mechanisms of invasion; however, models allowing analysis of shared pathways mediating invasion and therapy resistance are lacking. To evaluate therapy response associated with cancer cell invasion, we here used 3D invasion culture of tumor spheroids in 3D fibrillar collagen and applied Ethanol-Ethyl cinnamate (EtOH-ECi) based optical clearing to detect both spheroid core and invasion zone by subcellular-resolved 3D microscopy. When subjected to a single dose of irradiation (4 Gy), we detected significant cell survival in the invasion zone. By physical separation of the core and invasion zone, we identified differentially regulated genes preferentially engaged in invading cells controlling cell division, repair, and survival. This imaging-based 3D invasion culture may be useful for the analysis of complex therapy-response patterns in cancer cells in drug discovery and invasion-associated resistance development.
The online version contains supplementary material available at 10.1007/s44164-022-00040-x.
已开发出细胞外基质（ECM）中癌细胞的3D体外培养模型，用于研究药物靶向性和耐药性，或者用于研究侵袭机制；然而，缺乏能够分析介导侵袭和治疗耐药性的共同途径的模型。为了评估与癌细胞侵袭相关的治疗反应，我们在此使用了肿瘤球体在3D纤维状胶原蛋白中的3D侵袭培养，并应用基于乙醇-肉桂酸乙酯（EtOH-ECi）的光学透明化技术，通过亚细胞分辨3D显微镜检测球体核心和侵袭区域。当给予单剂量照射（4 Gy）时，我们在侵袭区域检测到显著的细胞存活。通过对核心和侵袭区域进行物理分离，我们鉴定出差异调节的基因，这些基因优先参与控制细胞分裂、修复和存活的侵袭细胞。这种基于成像的3D侵袭培养可能有助于分析药物发现中癌细胞的复杂治疗反应模式以及与侵袭相关的耐药性发展。
在线版本包含可在10.1007/s44164-022-00040-x获取的补充材料。
