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一个基于流细胞计的管道,用于快速量化C2C12细胞分化

Bianca Parisi¹, Maxime Sünnen¹, Rohan Chippalkatti¹

¹Cancer Cell Biology and Drug Discovery Group, Department of Life Sciences and Medicine, University of Luxembourg, 4362 Esch-sur-Alzette, Luxembourg.

|October 11, 2023

概括

这项研究引入了一种流细胞计法,用于测量C2C12细胞分化,使用髓重链标记物. FlowFate R脚本自动化了这种分析,使得对基因对差异化的影响进行研究.

科学领域:

细胞生物学细胞生物学
生物技术是生物技术.
分子生物学分子生物学

背景情况:

C2C12细胞系是广泛使用的体外模型,用于研究骨肌肉分化.
精确量化细胞分化对于理解肌肉发育和疾病至关重要.
评估差异化的现有方法可能是劳动密集型和主观的.

研究的目的:

开发和验证基于流细胞计的管道,用于定量评估C2C12细胞分化.
引入一种自动化分析工具,即FlowFate R脚本,用于处理流细胞计数据.
证明管道和脚本在评估特定基因对差异化的剂量依赖影响方面的实用性.

主要方法:

建立了C2C12细胞培养的详细方案,包括播种,转染,药物治疗和分化诱导.
利用髓重链表达作为分化髓管的标记物.
采用流细胞计量来获得高通量细胞分化数据,并开发了用于自动分析的FlowFate R脚本.

主要成果:

成功开发了一种强大的流细胞计管道,能够根据肌重链表达量化C2C12细胞分化.
FlowFate R脚本提供了流细胞计数据的自动化和可重复分析.
证明了使用开发的管道和脚本评估GFP标记基因对C2C12细胞分化的剂量依赖影响的能力.

结论:

本文介绍的流细胞计管道为研究C2C12细胞分化提供了一种定量和高效的方法.
FlowFate R脚本显著简化了差异化数据的分析,促进了高通量选和机制研究.
这种方法为研究人员研究调节骨肌肉分化分子机制提供了有价值的工具.

关键词:

癌症癌症癌症癌症细胞生物学细胞生物学分子生物学分子生物学

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相关概念视频

Flow Cytometry

Flow Cytometry

The development of flow cytometry techniques began in 1934 with initial attempts by Andrew Moldavan, a bacteriologist who counted the cells in a flowing capillary system. Moldavan pumped cells through a capillary tube focused under a microscope for visualization. The invention of photometry allowed the measurement of differentially-stained cells, and Louis Kamentsky developed the first multiparameter flow cytometer in 1965 to identify and count the cancer cells in cervical tissue specimens.
In...