英文原题:AI-based analysis of label-free live cell imaging of T-cell mediated tumor killing assay enables competitive and robust hit calling.
AI-based analysis of label-free live cell imaging of T-cell mediated tumor killing assay enables competitive and robust hit calling.
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利用T细胞受体(TCR)T细胞疗法和双特异性抗体(BsAb)等免疫细胞疗法发现及优化个体化癌症治疗时,必须通过免疫介导杀伤实验确认候选疗法具有稳健的功能活性。在这类实验中,研究者常使用自动显微镜对多种细胞系与患者来源原代癌细胞的共培养体系进行活细胞成像。传统方法使用荧光染料或特异表达的核蛋白进行标记,再通过依赖细胞分割的经典图像分析进行评估。
因此,这些方法易受光毒性和荧光漂白等伪影影响;细胞视觉表型随时间变化也会导致分割不准确,而且针对不同人体组织类型或供者的实验,分析参数需要不断调整。
本研究提出一种新方法,将明场图像与无需人工干预、可扩展的人工智能(AI)分析流程结合,不需要任何荧光标记。研究者将该流程用于T细胞介导的杀伤实验,并与当前基于荧光图像、半人工细胞分割的分析方法进行基准比较。新流程适用于表型多样的癌细胞,因省去人工调整步骤而效率更高,且结果一致性相当。研究认为,该AI分析流程有望大幅简化T细胞介导的活细胞杀伤实验,免除标记步骤,并直接分析明场图像,避免传统分割方法对标记图像进行耗时且困难的分析。
For the discovery and optimization of personalized cancer treatments using immune cell therapeutics, such as T-cell receptor (TCR-T) therapy and bispecific antibodies (BsAbs), robust functional activity of candidates must be confirmed in immune-mediated killing assays. In these assays, co-cultures of several cell lines and patient-derived primary cancer cells often are imaged live using automated microscopy.
Conventionally, such assays use fluorescent dyes or specifically expressed nuclear proteins for labeling, followed by classical image analysis reliant on cell segmentation. They are therefore subject to artifacts like phototoxicity and bleaching, inaccurate segmentation due to the typical variations in visual phenotype with time as well as requiring the constant adaptation of analysis parameters for experiments across different human tissue types or donors.
Here we present a new approach utilizing brightfield images in combination with a hands-free, scalable artificial intelligence (AI)-based analysis workflow, requiring no fluorescent markers at all.
We have applied this new workflow to a T-cell mediated killing assay and benchmarked it against current semi-manual, cell segmentation-based analysis of fluorescent images.
We found that the new workflow performs well on phenotypically diverse cancer cells, with greater efficiency though elimination of manual adjustment steps, and produces results of equivalent consistency.
We conclude that this AI-based analysis workflow has the potential to substantially simplify T-cell mediated live cell killing assays, eliminating the need for labeling, and allows their efficient analysis, operating on brightfield images and thus avoiding time-consuming and difficult analysis of labeled images using classical segmentation-based analysis.
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