Cristina Bucchi, Josefa Baeza, Jaime Guarda, Ana Bucchi, Paulina Martínez-Rodríguez
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引用次数: 0
摘要
背景:细胞生物学中通常使用划痕试验来评估细胞迁移;然而,这并不是一种标准化的方法,它产生的间隙尺寸变化很大。我们设计了一种可打印的装置,由单个缠绕工具和导向器组成,并比较了我们的装置和传统方法产生的间隙。该装置可用标准 3D 打印机打印。细胞被播种在 24 孔板上。细胞达到完全融合后,使用传统方法(用移液管吸头进行划痕试验)、移液管吸头和该装置的导向器或单个缠绕工具和导向器产生间隙。在光学显微镜下观察缝隙长达 48 小时并进行分析:结果表明,传统方法产生的间隙不规则、不平直,与其他组相比,细胞迁移率最差。伤痕工具在孔表面产生刮痕:结论:在划痕试验中,导板和移液管吸头的效果最好:意义:在划痕实验中使用导向器和移液器吸头可以进行可重复的细胞迁移实验。
A cost-effective tool to standardize the scratch assay for cell migration
Background
The scratch assay is commonly used in cell biology to evaluate cell migration; however, it is not a standardized method; it produces highly variable gap dimensions. We design a printable device, comprising a single wounding tool and a guide, and compared the gap produced by our device and the traditional method. The deviceis printable in a standard 3D printer. Cells were seeded on a 24-well plate. After reaching full confluency, a gap was created using the traditional method (scratch assay with a pipette tip), a pipette tip and the guide of the device, or the single wounding tool and the guide. The gaps were observed for up to 48 h under a light microscope and analyzed.
Results
The results show that the traditional method produces irregular and not straight gaps, and had the worst cell migration rates compared to the other groups. The wounding tool produced scrape signs at the well surface.
Conclusion
The guide and pipette tip delivered the best results for the scratch assay.
Significance
The use of the guide and the pipette tip for the scratch assay allows allows to perform reproducible cell migration experiments.
期刊介绍:
The journal publishes original research articles and reviews on all aspects of cellular, molecular and structural biology, developmental biology, cell physiology and evolution. It will publish articles or reviews contributing to the understanding of the elementary biochemical and biophysical principles of live matter organization from the molecular, cellular and tissues scales and organisms.
This includes contributions directed towards understanding biochemical and biophysical mechanisms, structure-function relationships with respect to basic cell and tissue functions, development, development/evolution relationship, morphogenesis, stem cell biology, cell biology of disease, plant cell biology, as well as contributions directed toward understanding integrated processes at the organelles, cell and tissue levels. Contributions using approaches such as high resolution imaging, live imaging, quantitative cell biology and integrated biology; as well as those using innovative genetic and epigenetic technologies, ex-vivo tissue engineering, cellular, tissue and integrated functional analysis, and quantitative biology and modeling to demonstrate original biological principles are encouraged.