Close David A, Camarco Daniel P, Shan Feng, Kochanek Stanton J, Johnston Paul A
Department of Pharmaceutical Sciences, University of Pittsburgh, Pittsburgh, PA, 15261, USA.
Department of Pharmaceutical Sciences, School of Pharmacy, University of Pittsburgh, Room 586 Salk Hall, 3501 Terrace Street, Pittsburgh, PA, 15261, USA.
Methods Mol Biol. 2018;1683:355-369. doi: 10.1007/978-1-4939-7357-6_20.
The poor success rate of cancer drug discovery has prompted efforts to develop more physiologically relevant cellular models for early preclinical cancer lead discovery assays. For solid tumors, this would dictate the implementation of three-dimensional (3D) tumor models that more accurately recapitulate human solid tumor architecture and biology. A number of anchorage-dependent and anchorage-independent in vitro 3D cancer models have been developed together with homogeneous assay methods and high content imaging approaches to assess tumor spheroid morphology, growth, and viability. However, several significant technical challenges have restricted the implementation of some 3D models in HTS. We describe a method that uses 384-well U-bottomed ultra-low attachment (ULA) microplates to produce head and neck tumor spheroids for cancer drug discovery assays. The production of multicellular head and neck cancer spheroids in 384-well ULA-plates occurs in situ, does not impose an inordinate tissue culture burden for HTS, is readily compatible with automation and homogeneous assay detection methods, and produces high-quality uniform-sized spheroids that can be utilized in cancer drug cytotoxicity assays within days rather than weeks.
癌症药物研发的成功率较低，这促使人们努力开发更具生理相关性的细胞模型，用于临床前癌症早期先导物发现试验。对于实体瘤而言，这意味着要采用三维（3D）肿瘤模型，该模型能更准确地再现人类实体瘤的结构和生物学特性。现已开发出多种依赖贴壁和不依赖贴壁的体外3D癌症模型，以及用于评估肿瘤球体形态、生长和活力的均质检测方法和高内涵成像方法。然而，一些重大的技术挑战限制了某些3D模型在高通量筛选（HTS）中的应用。我们描述了一种方法，该方法使用384孔U型底超低吸附（ULA）微孔板来生成用于癌症药物发现试验的头颈部肿瘤球体。在384孔ULA板中生成多细胞头颈部癌球体是原位进行的，不会给HTS带来过度的组织培养负担，很容易与自动化和均质检测方法兼容，并能生成高质量、大小均匀的球体，这些球体可在数天而非数周内用于癌症药物细胞毒性试验。
