Time-lapse image analysis for whole colony growth curves and daily distribution of the cell number per colony during the expansion of mesenchymal stem cells
Time-lapse image analysis for whole colony growth curves and daily distribution of the cell number per colony during the expansion of mesenchymal stem cells
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DOI：
10.1038/s41598-019-53383-z
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发表时间：
2019-11-14
影响因子：
4.6
通讯作者：
Sekiya, Ichiro
中科院分区：
文献类型：
--
作者：
Mizuno, Mitsuru;Katano, Hisako;Sekiya, Ichiro
来自滑膜的间充质干细胞（滑膜MSC）对于软骨和半月板再生治疗是有吸引力的。我们开发了一个软件程序，可以区分单个菌落，并使用延时图像自动计算每个菌落的细胞数。在这项研究中，我们调查了软件的有用性，并分析了培养的滑膜间充质干细胞的集落形成。获得了14天扩增的人滑膜MSC的延时图像数据。根据相差和核染色图像自动计数每个集落的细胞数（145个集落）。使用聚类分析对从第1天到第14天的菌落生长曲线（对于140个菌落）进行分类。第14天每个集落的细胞数分布与第1-14天的细胞数分布的相关性分析显示，第7天和第14天存在相关性。我们从相衬图像中获得了准确的细胞数计数。单个菌落生长曲线分为三个主要组和亚组。我们的图像分析软件有可能改善细胞增殖的评估，并促进使用MSC的成功临床应用。
Mesenchymal stem cells from the synovium (synovial MSCs) are attractive for cartilage and meniscus regeneration therapy. We developed a software program that can distinguish individual colonies and automatically count the cell number per colony using time-lapse images. In this study, we investigated the usefulness of the software and analyzed colony formation in cultured synovial MSCs. Time-lapse image data were obtained for 14-day-expanded human synovial MSCs. The cell number per colony (for 145 colonies) was automatically counted from phase-contrast and nuclear-stained images. Colony growth curves from day 1 to day 14 (for 140 colonies) were classified using cluster analysis. Correlation analysis of the distribution of the cell number per colony at 14 days versus that number at 1-14 days revealed a correlation at 7 and 14 days. We obtained accurate cell number counts from phase-contrast images. Individual colony growth curves were classified into three main groups and subgroups. Our image analysis software has the potential to improve the evaluation of cell proliferation and to facilitate successful clinical applications using MSCs.