Cell Dynamics iSRL, Bologna, Italy.
RAMSES Laboratory, IRCCS Istituto Ortopedico Rizzoli, Bologna, Italy.
PLoS One. 2021 Jun 8;16(6):e0252907. doi: 10.1371/journal.pone.0252907. eCollection 2021.
Three-dimensional (3D) culture systems like tumor spheroids represent useful in vitro models for drug screening and more broadly for cancer biology research, but the generation of uniform populations of spheroids remains challenging. The possibility to properly characterize spheroid properties would increase the reliability of these models. To address this issue different analysis were combined: i) a new device and relative analytical method for the accurate, simultaneous, and rapid measurement of mass density, weight, and size of spheroids, ii) confocal imaging, and iii) protein quantification, in a clinically relevant 3D model. The LoVo colon cancer cell line forming spheroids, treated with crizotinib (CZB) an ATP-competitive small-molecule inhibitor of the receptor tyrosine kinases, was employed to study and assess the correlation between biophysical and morphological parameters in both live and fixed cells. The new fluidic-based measurements allowed a robust phenotypical characterization of the spheroids structure, offering insights on the spheroids bulk and an accurate measurement of the tumor density. This analysis helps overcome the technical limits of the imaging that hardly penetrates the thickness of 3D structures. Accordingly, we were able to document that CZB treatment has an impact on mass density, which represents a key marker characterizing cancer cell treatment. Spheroid culture is the ultimate technology in drug discovery and the adoption of such precise measurement of the tumor characteristics can represent a key step forward for the accurate testing of treatment's potential in 3D in vitro models.
三维（3D）培养系统，如肿瘤球体，代表了用于药物筛选和更广泛的癌症生物学研究的有用的体外模型，但生成均匀的球体群体仍然具有挑战性。正确表征球体特性的可能性将提高这些模型的可靠性。为了解决这个问题，我们结合了不同的分析方法：i）一种新的设备和相关的分析方法，用于准确、同时和快速测量球体的质量密度、重量和大小，ii）共聚焦成像，以及 iii）在临床相关的 3D 模型中进行蛋白质定量。采用形成球体的 LoVo 结肠癌细胞系，用克唑替尼（CZB）处理，CZB 是受体酪氨酸激酶的一种 ATP 竞争性小分子抑制剂，用于研究和评估活细胞和固定细胞中生物物理和形态参数之间的相关性。新的基于流体的测量方法允许对球体结构进行稳健的表型特征分析，提供有关球体整体的见解，并对肿瘤密度进行准确测量。这种分析有助于克服成像技术的技术限制，这些限制很难穿透 3D 结构的厚度。因此，我们能够证明 CZB 治疗对质量密度有影响，这是表征癌细胞治疗的关键标志物。球体培养是药物发现的终极技术，采用这种精确测量肿瘤特性的方法可以代表在 3D 体外模型中准确测试治疗潜力的关键一步。
