A multi-inlet microfluidic device fabricated for in situ detection of multiple cytotoxicity endpoints
A multi-inlet microfluidic device fabricated for in situ detection of multiple cytotoxicity endpoints
复制标题
DOI：
10.1007/s13206-012-6107-6
复制
发表时间：
2012-03-20
期刊：
影响因子：
4.3
通讯作者：
Rhee, Seog Woo
中科院分区：
文献类型：
--
作者：
Mahto, Sanjeev Kumar;Rhee, Seog Woo
本工作描述了一种用于原位检测多个细胞毒性终点的多入口微流控装置的研制。该装置由上游微流控多入口模块(Mu MIM)和下游微流控细胞培养通道(Mu CCC)组成。通过MU MIM的入口处将设备与注射器泵集成在一起，主要实现了针对多个终点的多路细胞毒性检测。本实验装置用于BALB/3T3成纤维细胞的长期培养。此外，还用镉离子作为毒物对培养的细胞进行了毒性试验。同时检测单个细胞暴露于镉后的细胞毒性终点，如活性氧自由基(ROS)的原位检测、核结构和细胞形态。数据表明，产生ROS并显示核染色的细胞的百分比略高于仅产生ROS的细胞，但这一百分比明显高于仅显示核染色的细胞。结果表明，从多重细胞毒性同时观察获得的相关数据在为Cd诱导的毒性提供机制解释方面比在检查单一终点后获得的单独数据更有效。此外，这些结果表明，镉诱导细胞内ROS的形成可能伴随着细胞和细胞核形态的改变和膜完整性的丧失。该设备允许对细胞及其针对毒物的行为进行实时监测，并有助于快速进行细胞毒性测试。此外，我们预计，通过提高识别终点的灵敏度和准确性，所提供的设备可以非常有效地促进对细胞毒性的精确机制的理解。
This work describes the development of a multi-inlet microfluidic device for in situ detection of multiple cytotoxicity endpoints. The device consists of an upstream microfluidic multi-inlet module (mu MIM) and a downstream microfluidic cell culture channel (mu CCC). The integration of a device with syringe pumps via the inlets of mu MIM mainly enables the performance of multiplex cytotoxicity assays targeted on multiple endpoints. The device in the study was used for the long-term culturing of BALB/3T3 fibroblast cells. In addition, cadmium ion was used as a toxicant to induce the toxicity in the cultured cells. The cytotoxicity endpoints, such as the in situ detection of reactive oxygen species (ROS), nuclear structures and cell morphology, were examined and correlated simultaneously for every single cell after they were exposed to cadmium. The data indicated that the percentage of cells that had produced ROS and shown nuclear staining were slightly higher than the cells that had produced ROS only, but this percentage was significantly higher than the cells that had shown only nuclear staining. The results revealed that the obtained correlated data from simultaneous observations of multiplex cytotoxicity assays were more efficient in providing the mechanistic explanation for Cd-induced toxicity than the individual data that were derived after examining a single endpoint. In addition, the results suggested that the intracellular formation of ROS induced by cadmium might have been followed by the alterations in cell and nuclear morphology and loss of membrane integrity. The device allows for the real-time monitoring of cells and their behavior against a toxicant and facilitates the rapid performance of cytotoxicity testing. Furthermore, we anticipate that the presented device could be very effective in advancing the understanding of the precise mechanism of cytotoxicity by improving the sensitivity and accuracy in identifying endpoints.