Department of Chemistry "G. Ciamician", University of Bologna, 40126 Bologna, Italy; INBB, Istituto Nazionale di Biostrutture e Biosistemi, 00136 Rome, Italy.
Department of Chemistry "G. Ciamician", University of Bologna, 40126 Bologna, Italy.
Biosens Bioelectron. 2019 Jan 1;123:269-277. doi: 10.1016/j.bios.2018.09.012. Epub 2018 Sep 4.
Whole-cell biosensors present many advantages, including being able to monitor the toxicity and bioavailability of chemicals; cells grown in traditional 2D cultures, however, do not reproduce the complexity of in vivo physiology. In the last years, 3D cell-culture models have garnered great attention due to their capability to better mimic in vivo cellular responses to external stimuli, providing excellent model living organisms. In order to obtain a predictive, sensitive, and robust yet low-cost 3D cell biosensor, we developed a smartphone-based bioluminescent 3D cell biosensor platform for effect-based analysis. We exploited the Nuclear Factor-kappa B (NF-kB) signal transduction pathway, which is induced by several types of stressors and is involved in the regulation of cell-cycle/growth, inflammation, apoptosis, and immunity. The smartphone-based biosensor relies on immobilized HEK293 spheroids genetically engineered with powerful red- and green-emitting luciferases utilized as inflammation and viability reporters. It provides a limit of detection for Tumor Necrosis Factor (TNFα) of 0.15 ± 0.05 ng/mL and could be a useful tool to initially screen environmental samples or other compounds on-site, especially for additional more accurate chemical analyses.
全细胞生物传感器具有许多优点，包括能够监测化学物质的毒性和生物利用度；然而，在传统的 2D 培养中生长的细胞无法复制体内生理学的复杂性。在过去几年中，由于能够更好地模拟体内细胞对外部刺激的反应，3D 细胞培养模型引起了极大的关注，为研究生物体提供了良好的模型。为了获得一种具有预测性、敏感性、稳健性且成本低的 3D 细胞生物传感器，我们开发了一种基于智能手机的生物发光 3D 细胞生物传感器平台，用于基于效应的分析。我们利用核因子-κB（NF-κB）信号转导途径，该途径由多种应激源诱导，并参与细胞周期/生长、炎症、细胞凋亡和免疫的调节。基于智能手机的生物传感器依赖于固定化的 HEK293 球体，这些球体经过基因工程改造，具有强大的红色和绿色荧光素酶，可用作炎症和活力报告器。它对肿瘤坏死因子 (TNFα) 的检测限为 0.15±0.05ng/mL，可作为初步筛选环境样品或其他化合物的有用工具，特别是用于现场进行其他更准确的化学分析。
