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Fast Adipogenesis Tracking System (FATS)—a robust high-throughput automation-ready adipogenesis quantification technique

机译:快速脂肪生成跟踪系统(FATS)-强大的高通量自动化就绪的脂肪生成量化技术

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摘要

Adipogenesis is essential in in vitro experimentation to assess differentiation capability of stem cells, and therefore, its accurate measurement is important. Quantitative analysis of adipogenic levels, however, is challenging and often susceptible to errors due to non-specific reading or manual estimation by observers. To this end, we developed a novel adipocyte quantification algorithm, named Fast Adipogenesis Tracking System (FATS), based on computer vision libraries. The FATS algorithm is versatile and capable of accurately detecting and quantifying percentage of cells undergoing adipogenic and browning differentiation even under difficult conditions such as the presence of large cell clumps or high cell densities. The algorithm was tested on various cell lines including 3T3-L1 cells, adipose-derived mesenchymal stem cells (ASCs), and induced pluripotent stem cell (iPSC)-derived cells. The FATS algorithm is particularly useful for adipogenic measurement of embryoid bodies derived from pluripotent stem cells and was capable of accurately distinguishing adipogenic cells from false-positive stains. We then demonstrate the effectiveness of the FATS algorithm for screening of nuclear receptor ligands that affect adipogenesis in the high-throughput manner. Together, the FATS offer a universal and automated image-based method to quantify adipocyte differentiation of different cell lines in both standard and high-throughput workflows.Electronic supplementary materialThe online version of this article (10.1186/s13287-019-1141-0) contains supplementary material, which is available to authorized users.
机译:在体外实验中,成脂作用对于评估干细胞的分化能力至关重要,因此,其精确测量非常重要。但是,对脂肪形成水平的定量分析具有挑战性,并且由于非特异性读数或观察者的手动估计,常常容易出错。为此,我们基于计算机视觉库开发了一种新颖的脂肪细胞定量算法,称为快速脂肪生成跟踪系统(FATS)。 FATS算法用途广泛,即使在困难条件下(例如,存在大细胞团块或高细胞密度),也能准确检测和量化经历成脂和褐变的细胞百分比。在各种细胞系上测试了该算法,包括3T3-L1细胞,脂肪来源的间充质干细胞(ASC)和诱导多能干细胞(iPSC)来源的细胞。 FATS算法对于源自多能干细胞的胚状体的成脂测量特别有用,并且能够准确地将成脂细胞与假阳性染色区分开。然后,我们证明了FATS算法可用于筛选以高通量方式影响脂肪生成的核受体配体。 FATS共同提供了一种基于图像的通用且自动化的方法,可以定量标准和高通量工作流程中不同细胞系的脂肪细胞分化。电子补充材料本文的在线版本(10.1186 / s13287-019-1141-0)包含补充材料,授权用户可以使用。

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