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Supported gel slab scaffolds as a three-dimensional cell-based assay platform
Analyst ( IF 3.6 ) Pub Date : 2024-07-22 , DOI: 10.1039/d4an00691g Zachary R Sitte 1 , Elizabeth E Karlsson 1 , Tyler S Larson 1 , Haolin Li 2 , Haibo Zhou 2, 3 , Matthew R Lockett 1, 4
Analyst ( IF 3.6 ) Pub Date : 2024-07-22 , DOI: 10.1039/d4an00691g Zachary R Sitte 1 , Elizabeth E Karlsson 1 , Tyler S Larson 1 , Haolin Li 2 , Haibo Zhou 2, 3 , Matthew R Lockett 1, 4
Affiliation
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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 in vivo 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.
中文翻译：
支持凝胶板支架作为三维细胞分析平台
基于细胞的检测在药物发现流程中受到严重依赖，可快速将大型化合物库与可管理数量的候选药物进行配对，以进行进一步的表征和评估。将细胞沉积在孔板底部的单层培养物是许多此类筛选的主力，尽管不断有证据表明它们无法预测体内反应。三维 (3D) 培养平台可以生成比单层细胞更具代表性的细胞表型的组织样环境，但事实证明，将其纳入已开发的工作流程中具有挑战性。基于支架的方法是一种生成组织样环境的易处理方法，支持载有细胞的凝胶，其制备类似于将细胞沉积在孔板中。在这里，我们描述了由市售材料、粘合剂喷雾和激光切割机制备的支撑凝胶板（SGS）支架。这些含有细胞的支架可以很容易地装入孔板中，提供与当前液体处理器和分析仪器兼容的格式。该支架能够评估单个或堆叠结构中的细胞反应，其中含有富含细胞外基质的微环境。通过一系列的演示，我们强调了易于组装的 SGS 支架在量化细胞反应方面的实用性。这些读数包括共焦显微镜、量化类似 Transwell 和堆叠格式的细胞侵袭、生成能够提供药物反应空间分辨图的多层按需结构，以及在筛选应用中识别潜在的化疗。
更新日期：2024-07-22
中文翻译：
支持凝胶板支架作为三维细胞分析平台
基于细胞的检测在药物发现流程中受到严重依赖，可快速将大型化合物库与可管理数量的候选药物进行配对，以进行进一步的表征和评估。将细胞沉积在孔板底部的单层培养物是许多此类筛选的主力，尽管不断有证据表明它们无法预测体内反应。三维 (3D) 培养平台可以生成比单层细胞更具代表性的细胞表型的组织样环境，但事实证明，将其纳入已开发的工作流程中具有挑战性。基于支架的方法是一种生成组织样环境的易处理方法，支持载有细胞的凝胶，其制备类似于将细胞沉积在孔板中。在这里，我们描述了由市售材料、粘合剂喷雾和激光切割机制备的支撑凝胶板（SGS）支架。这些含有细胞的支架可以很容易地装入孔板中，提供与当前液体处理器和分析仪器兼容的格式。该支架能够评估单个或堆叠结构中的细胞反应，其中含有富含细胞外基质的微环境。通过一系列的演示，我们强调了易于组装的 SGS 支架在量化细胞反应方面的实用性。这些读数包括共焦显微镜、量化类似 Transwell 和堆叠格式的细胞侵袭、生成能够提供药物反应空间分辨图的多层按需结构，以及在筛选应用中识别潜在的化疗。
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