Evaluating Cell Processes, Quality, and Biomarkers in Pluripotent Stem Cells Using Video Bioinformatics.
Evaluating Cell Processes, Quality, and Biomarkers in Pluripotent Stem Cells Using Video Bioinformatics.
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DOI：
10.1371/journal.pone.0148642
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发表时间：
2016
期刊：
影响因子：
3.7
通讯作者：
Talbot P
中科院分区：
文献类型：
--
作者：
Zahedi A;On V;Lin SC;Bays BC;Omaiye E;Bhanu B;Talbot P
在从事基础研究、再生疗法和毒理学研究的干细胞实验室中，对质量控制工具有基本的需求。这些工具需要在体外传代、扩增、维持和分化期间评估细胞过程和质量的自动化方法。在本文中，一个公正的，自动化的高内容分析工具，StemCellQC，提出了非侵入性提取信息的细胞质量和细胞过程的时间推移相衬视频。在健康、不健康和垂死的人胚胎干细胞（hESC）集落中分析了二十四（24）个形态学和动态特征，以鉴定每组中受影响的那些特征。健康组与不健康/垂死组的多个特征不同，这些特征与生长，运动和死亡有关。发现了生物标记物，它们在人工观察之前就预测了细胞过程。StemCellQC通过非侵入性测量和跟踪48小时内的动态和形态特征，以96%的准确率区分健康和不健康/垂死的hESC集落。StemCellQC可以监测细胞过程的变化，并预测多能干细胞集落的质量。该工具包减少了跟踪多个多能干细胞集落所需的时间和资源，并消除了由于人为偏见而导致的处理错误和错误分类。StemCellQC在受影响的功能不直观或预期的情况下提供用户指定和分类器确定的分析。视频分析算法允许使用自动检测分析评估生物现象，这可以帮助维持干细胞质量和/或监测细胞过程变化至关重要的设施。在未来，StemCellQC可以扩展到包括其他功能，细胞类型，治疗和分化细胞。
There is a foundational need for quality control tools in stem cell laboratories engaged in basic research, regenerative therapies, and toxicological studies. These tools require automated methods for evaluating cell processes and quality during in vitro passaging, expansion, maintenance, and differentiation. In this paper, an unbiased, automated high-content profiling toolkit, StemCellQC, is presented that non-invasively extracts information on cell quality and cellular processes from time-lapse phase-contrast videos. Twenty four (24) morphological and dynamic features were analyzed in healthy, unhealthy, and dying human embryonic stem cell (hESC) colonies to identify those features that were affected in each group. Multiple features differed in the healthy versus unhealthy/dying groups, and these features were linked to growth, motility, and death. Biomarkers were discovered that predicted cell processes before they were detectable by manual observation. StemCellQC distinguished healthy and unhealthy/dying hESC colonies with 96% accuracy by non-invasively measuring and tracking dynamic and morphological features over 48 hours. Changes in cellular processes can be monitored by StemCellQC and predictions can be made about the quality of pluripotent stem cell colonies. This toolkit reduced the time and resources required to track multiple pluripotent stem cell colonies and eliminated handling errors and false classifications due to human bias. StemCellQC provided both user-specified and classifier-determined analysis in cases where the affected features are not intuitive or anticipated. Video analysis algorithms allowed assessment of biological phenomena using automatic detection analysis, which can aid facilities where maintaining stem cell quality and/or monitoring changes in cellular processes are essential. In the future StemCellQC can be expanded to include other features, cell types, treatments, and differentiating cells.