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A Modified Limiting Dilution Method for Monoclonal Stable Cell Line Selection Using a Real-Time Fluorescence Imaging System: A Practical Workflow and Advanced Applications

机译:使用实时荧光成像系统的单克隆稳定细胞系选择改进的限制稀释方法:实用的工作流程和高级应用

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摘要

Stable cell lines are widely used in laboratory research and pharmaceutical industry. They are mainly applied in recombinant protein and antibody productions, gene function studies, drug screens, toxicity assessments, and for cancer therapy investigation. There are two types of cell lines, polyclonal and monoclonal origin, that differ regarding their homogeneity and heterogeneity. Generating a high-quality stable cell line, which can grow continuously and carry a stable genetic modification without alteration is very important for most studies, because polyclonal cell lines of multicellular origin can be highly variable and unstable and lead to inconclusive experimental results. The most commonly used technologies of single cell originate monoclonal stable cell isolation in laboratory are fluorescence-activated cell sorting (FACS) sorting and limiting dilution cloning. Here, we describe a modified limiting dilution method of monoclonal stable cell line selection using the real-time fluorescence imaging system IncuCyte®S3.
机译:稳定的细胞系广泛用于实验室研究和制药行业。它们主要应用于重组蛋白和抗体制作,基因函数研究,药物筛网,毒性评估以及癌症治疗调查。有两种类型的细胞系,多克隆和单克隆来源,其均匀性和异质性不同。产生高质量的稳定细胞系,其可以连续生长并携带稳定的遗传修饰而不改变对于大多数研究非常重要,因为多细胞来源的多克隆细胞系可以高度可变和不稳定,并且导致不确定的实验结果。在实验室中,单胞细胞的最常用技术源自单克隆稳定的细胞分离是荧光激活细胞分选(FACS)分选和限制稀释克隆。在这里,我们描述了使用实时荧光成像系统INCUCYTES3的单克隆稳定细胞系选择的修饰限制稀释方法。

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