Department of Urology, School of Medicine, Stony Brook University, Stony Brook, NY 11794, USA.
Department of Pathology, School of Medicine, Stony Brook University, Stony Brook, NY 11794, USA.
Biotechniques. 2020 Sep;69(3):193-199. doi: 10.2144/btn-2020-0068. Epub 2020 Jul 13.
3D cancer cell models are suitable for drug evaluation because they more precisely mimic tissue architecture than 2D cultures. To study cytotoxicity of anticancer agents, the most sensitive CellTiter-Glo 3D assay is used. However, this is an end point assay, so it is not possible to consider the variance of the starting material amount in the final reading. It is difficult to maintain an even plating density of 3D organoids for cytotoxicity analysis. We present a simple, 3D bladder cancer culture that can be maintained, cryopreserved and used for molecular and drug response studies. We applied a simple modification of the drug response assay for 3D cultures by measuring the background signal with the CellTiter Blue assay before drug application.
3D 癌细胞模型更适合药物评估,因为它们比 2D 培养物更能准确模拟组织结构。为了研究抗癌药物的细胞毒性,通常使用最敏感的 CellTiter-Glo 3D 检测法。然而,这是一种终点检测法,因此无法考虑最终读数中起始材料数量的变化。很难维持 3D 类器官用于细胞毒性分析的均匀接种密度。我们提出了一种简单的膀胱癌培养方法,可进行培养、冷冻保存并用于分子和药物反应研究。我们在药物应用前通过 CellTiter Blue 检测法测量背景信号,对 3D 培养物的药物反应检测法进行了简单的修改。