A high-content platform to characterise human induced pluripotent stem cell lines.
A high-content platform to characterise human induced pluripotent stem cell lines.
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DOI：
10.1016/j.ymeth.2015.11.012
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发表时间：
2016-03-01
期刊：
影响因子：
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通讯作者：
Danovi D
中科院分区：
文献类型：
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作者：
Leha A;Moens N;Meleckyte R;Culley OJ;Gervasio MK;Kerz M;Reimer A;Cain SA;Streeter I;Folarin A;Stegle O;Kielty CM;HipSci Consortium;Durbin R;Watt FM;Danovi D
IPSC显示出内部/内部/捐赠者的可变性阻碍了特征的确定。HipSci从数百名个人捐赠者那里产生、储存和提供IPSC。在96孔检测中，IPSCs对不同浓度的人血浆纤维连接蛋白有反应。表型特征：细胞数量、增殖、形态和细胞间黏附。所描述的方法可以为疾病建模和其他细胞类型量身定做。诱导多能干细胞(IPSCs)为未来的细胞治疗以及研究人类发育、疾病模型和发现治疗方法提供了宝贵的机会。为了实现IPSCs的潜力，对从大量健康和患病个体产生的细胞进行全面表征是至关重要的。人类IPSC倡议(HipSci)正在评估一大批细胞系，以确定细胞表型，剖析线内和线内以及供体的可变性，并确定其关键决定因素。在这里，我们报告了一个高含量的平台的建立，用于人类IPSC系的表型分析。在描述的实验中，细胞被分离并作为单个细胞种植在96孔板上，上面覆盖着三种不同浓度的纤维连接蛋白。这种方法可以评估细胞的数量、增殖、形态和细胞间的黏附。总而言之，我们的策略提供了复杂细胞群体内表型多样性的可靠量化，便于未来识别变异的遗传、生物和技术决定因素。所描述的方法可用于对来自多个捐赠者的IPSC进行基准测试，并创建可随时为疾病建模和药物发现量身定做的新平台。
iPSCs show inter/intra-line/donor-variability hampering characterisation. HipSci generates, banks and provides iPSCs from hundreds of individual donors. iPSCs respond to different human plasma fibronectin concentrations on 96-well assays. Phenotypic features: cell number, proliferation, morphology and intercellular adhesion. The methodologies described can be tailored for disease-modelling and other cell types. Induced pluripotent stem cells (iPSCs) provide invaluable opportunities for future cell therapies as well as for studying human development, modelling diseases and discovering therapeutics. In order to realise the potential of iPSCs, it is crucial to comprehensively characterise cells generated from large cohorts of healthy and diseased individuals. The human iPSC initiative (HipSci) is assessing a large panel of cell lines to define cell phenotypes, dissect inter- and intra-line and donor variability and identify its key determinant components. Here we report the establishment of a high-content platform for phenotypic analysis of human iPSC lines. In the described assay, cells are dissociated and seeded as single cells onto 96-well plates coated with fibronectin at three different concentrations. This method allows assessment of cell number, proliferation, morphology and intercellular adhesion. Altogether, our strategy delivers robust quantification of phenotypic diversity within complex cell populations facilitating future identification of the genetic, biological and technical determinants of variance. Approaches such as the one described can be used to benchmark iPSCs from multiple donors and create novel platforms that can readily be tailored for disease modelling and drug discovery.