Live-cell imaging of subcellular structures for quantitative evaluation of pluripotent stem cells
Live-cell imaging of subcellular structures for quantitative evaluation of pluripotent stem cells
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DOI：
10.1038/s41598-018-37779-x
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发表时间：
2019-02-11
影响因子：
4.6
通讯作者：
Hisatake, Koji
中科院分区：
文献类型：
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作者：
Nishimura, Ken;Ishiwata, Hiroshi;Hisatake, Koji
多能干细胞具有不同程度的多能性，在将其应用于再生医学之前，有必要选择具有高多能性的多能干细胞。然而，PSCs的质量控制过程是昂贵和耗时的，开发廉价和省力的选择方法将PSCs转化为临床应用是至关重要的。在这里，我们开发了一种成像系统，称为相位分布(PD)成像系统，它可以定量显示未染色和未标记细胞的亚细胞结构。PD图像及其派生的PD指数反映了线粒体的含量，使定量评估体细胞重新编程和PSC分化的程度成为可能。此外，PD指数允许将PSC克隆无偏见地分组为具有高或低多能性的克隆，而不需要借助侵入性方法。最后，PD成像系统产生了PSC集落的三维图像，为评估PSC的多能性提供了进一步的标准。因此，在更严格的质量控制过程之前，PD成像系统可用于筛选具有潜在高多能性的活体PSCs。
Pluripotent stem cells (PSCs) have various degrees of pluripotency, which necessitates selection of PSCs with high pluripotency before their application to regenerative medicine. However, the quality control processes for PSCs are costly and time-consuming, and it is essential to develop inexpensive and less laborious selection methods for translation of PSCs into clinical applications. Here we developed an imaging system, termed Phase Distribution (PD) imaging system, which visualizes subcellular structures quantitatively in unstained and unlabeled cells. The PD image and its derived PD index reflected the mitochondrial content, enabling quantitative evaluation of the degrees of somatic cell reprogramming and PSC differentiation. Moreover, the PD index allowed unbiased grouping of PSC colonies into those with high or low pluripotency without the aid of invasive methods. Finally, the PD imaging system produced three-dimensional images of PSC colonies, providing further criteria to evaluate pluripotency of PSCs. Thus, the PD imaging system may be utilized for screening of live PSCs with potentially high pluripotency prior to more rigorous quality control processes.