Department of Chemistry, University of North Carolina at Chapel Hill, 125 South Road, Chapel Hill, NC 27599-3290, USA.
Department of Biostatistics, University of North Carolina at Chapel Hill, 135 Dauer Drive, Chapel Hill, NC 27599-7400, USA.
Analyst. 2024 Sep 9;149(18):4653-4662. doi: 10.1039/d4an00691g.
Cell-based assays are heavily relied on in the drug discovery pipeline, quickly pairing down large compound libraries to a manageable number of drug candidates for further characterization and evaluation. Monolayer cultures in which cells are deposited onto the bottom of well plates are the workhorse of many of these screens despite continued evidence of their inability to predict responses. Three-dimensional (3D) culture platforms can generate tissue-like environments with more representative cellular phenotypes than monolayers but have proven challenging to incorporate into already-developed workflows. Scaffold-based approaches are a tractable means of generating tissue-like environments, supporting cell-laden gels whose preparation is analogous to depositing cells in a well plate. Here, we describe supported gel slab (SGS) scaffolds prepared from commercially available materials, an adhesive spray, and a laser cutter. These cell-containing scaffolds can readily fit into well plates, providing a format compatible with current liquid handlers and analytical instrumentation. The scaffolds enable the evaluation of cellular responses in individual or stacked structures, which contain extracellular matrix-rich microenvironments. With a series of demonstrations, we highlight the utility of the readily assembled SGS scaffolds to quantify cellular responses. These readouts include confocal microscopy, quantifying cellular invasion in Transwell-like and stacked formats, generating multilayered spheroid-on-demand structures capable of providing spatially resolved maps of drug responses, and identifying potential chemotherapies in a screening application.
基于细胞的测定在药物发现管道中被广泛应用，能够迅速将大量化合物库筛选到可管理数量的药物候选物，以进行进一步的特征描述和评估。尽管有持续的证据表明单层培养无法预测反应，但单层培养仍然是许多这些筛选的主力。三维（3D）培养平台可以生成更具代表性的细胞表型的类似组织的环境，但已证明难以将其纳入已开发的工作流程中。支架基方法是生成类似组织的环境的可行手段，支持细胞负载的凝胶，其制备类似于在孔板中沉积细胞。在这里，我们描述了由市售材料、粘性喷雾和激光切割机制备的支撑凝胶片（SGS）支架。这些含有细胞的支架可以轻松地放入孔板中，为当前的液体处理和分析仪器提供了兼容的格式。支架允许在单个或堆叠结构中评估细胞反应，这些结构包含富含细胞外基质的微环境。通过一系列演示，我们突出了易于组装的 SGS 支架在量化细胞反应方面的实用性。这些读数包括共聚焦显微镜，定量 Transwell 样和堆叠格式中的细胞侵袭，生成按需多层球体结构，能够提供药物反应的空间分辨图谱，以及在筛选应用中识别潜在的化疗药物。
