A Unidirectional 96-Well Fluidic Culture Platform for Upstream Cell Dosing with Subsequent Downstream Nonlinear and Ascending Exposure Gradients for Real-Time and Cell-Based Toxicity Screening Environments.
A Unidirectional 96-Well Fluidic Culture Platform for Upstream Cell Dosing with Subsequent Downstream Nonlinear and Ascending Exposure Gradients for Real-Time and Cell-Based Toxicity Screening Environments.
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用于上游细胞给药的单向 96 孔流体培养平台，以及随后的下游非线性和上升曝光梯度，用于实时和基于细胞的毒性筛查环境。
DOI：
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发表时间：
2021
影响因子：
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通讯作者：
R. McClelland
中科院分区：
文献类型：
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作者：
Bincy A. John;D. J. Sloan;Timothy Jensen;S. Ramaiahgari;P. End;Gabrielle E. Resh;R. McClelland
简介：由于创造更好地模拟体内模型的体外筛选工具的重要性，对于药物或毒物的暴露反应，可重复性和适应性的培养平台必须演变为复制人类器官系统固有功能的方法。阶梯瀑布(Ss瀑布)流体培养系统是一种单向、多井、重力驱动的细胞培养系统，每行有微通道连接12个井(8行重复)。材料和方法：该结构允许基质、母体和代谢物化合物的单向流动，以及相连的培养井之间的细胞信号传递，同时作为级联流动和离散的非线性给药装置运行。Ss瀑布中的初始细胞接种模仿了传统的静态平板方案，但之后在受控流动和浓度随时间暴露的环境中发挥作用。结果：为了研究微流控系统在预测药物疗效和毒性方面的应用，我们首先描述了一次性剂量和梯度曝光平台的设计、制造和表征。我们首先对设备的各种特征进行详细的表征，包括低非特异性结合、润湿性、与多种细胞类型的生物兼容性、井内和井间流动，以及添加到平台中的剂量化合物的高效自动混合特性。讨论：我们使用顺序测试中的一个例子来演示该设备的实用性--在大范围的可诱导暴露中筛选药物毒性和有效性，0 → IC100，具有实时评估的特点。结论：与传统/费力的人工药物稀释和/或昂贵的机器人技术相比，集成的自动梯度技术--带阶梯通道的重力流动--为最终用户提供了一种简单、快速的替代方案，可以评估新化合物实体(如制药、环境、农业、化妆品)的广泛毒性。
Introduction: Because of the importance to create in vitro screening tools that better mimic in vivo models, for exposure responses to drugs or toxicants, reproducible and adaptable culture platforms must evolve as approaches to replicate functions that are native to human organ systems. The Stairstep Waterfall (SsWaterfall) Fluidic Culture System is a unidirectional, multiwell, gravity-driven, cell culture system with micro-channels connecting 12 wells in each row (8-row replicates). Materials and Methods: The construct allows for the one-way flow of medium, parent and metabolite compounds, and the cellular signaling between connected culture wells while simultaneously operating as a cascading flow and discretized nonlinear dosing device. Initial cell seeding in SsWaterfall mimics traditional static plate protocols but thereafter functions with controlled flow and ramping concentration versus time exposure environments. Results: To investigate the utility of a microfluidic system for predicting drug efficacy and toxicity, we first delineate device design, fabrication, and characterization of a disposable dosing and gradient-exposure platform. We start with detailed characterizations by demarcating various features of the device, including low nonspecific binding, wettability, biocompatibility with multiple cell types, intra-well and inter-well flow, and efficient auto-mixing properties of dose compounds added into the platform. Discussion: We demonstrate the device utility using an example in sequential testing-screening drug toxicity and efficacy across wide-ranging inducible exposures, 0 → IC100, featuring real-time assessments. Conclusion: The integrated auto-gradient technology, gravity flow with stairstep pathways, offers end-users an easy and quick alternative to evaluate broad-ranging toxicity of new compound entities (e.g., pharmaceutical, environmental, agricultural, cosmetic) as opposed to traditional/arduous manual drug dilutions and/or expensive robotic technology.