Visualization of morphological categories of colonies for monitoring of effect on induced pluripotent stem cell culture status
Visualization of morphological categories of colonies for monitoring of effect on induced pluripotent stem cell culture status
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DOI：
10.1016/j.reth.2016.12.003
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发表时间：
2017-06-01
影响因子：
4.3
通讯作者：
Kato, Ryuji
中科院分区：
文献类型：
--
作者：
Nagasaka, Risako;Matsumoto, Megumi;Kato, Ryuji
从最近的进展来看，人们对大量生产用于各种应用的诱导多能干细胞（iPSC）的期望越来越高。对于这种类型的工业电池制造，强烈需要能够稳定生产效率的技术。由于目前的iPSC培养被精细的手工操作所覆盖，因此来自相同培养方案的所产生的细胞仍然存在质量差异。为了对iPSC的培养过程进行实时监控，并通过量化数据对iPSC的培养状态进行评估，本文介绍了基于图像的iPSC集落形态多样性可视化方法。我们设置了三种类型的实验来评估iPSC培养技术中可能干扰iPSC集落未分化状态的影响因素：1)技术差异的通道技能，（Exp. 2)饲养细胞制备中的技术差异，和（实验）3)维护技能的技术差异（中等更换频率，结合手动去除形态不规则的菌落）。通过实时显微图像测量所有存在的菌落，可视化培养容器中菌落形态的异质性变化。通过使用形态分类的曼哈顿图的这种可视化，可以比较技术技能之间的差异，以评估适当的细胞处理。(C)2017年，日本再生医学学会。制作和主办：Elsevier B.V.
From the recent advances, there are growing expectations toward the mass production of induced pluripotent stem cells (iPSCs) for varieties of applications. For such type of industrial cell manufacturing, the technology which can stabilize the production efficiency is strongly required. Since the present iPSC culture is covered by delicate manual operations, there are still quality differences in produced cells from same culture protocols. To monitor the culture process of iPSCs with the quantified data to evaluate the culture status, we here introduce image-based visualization method of morphological diversity of iPSC colonies. We have set three types of experiments to evaluate the influential factors in iPSC culture technique that may disturb the undifferentiation status of iPSC colonies: (Exp. 1) technical differences in passage skills, (Exp. 2) technical differences in feeder cell preparation, and (Exp. 3) technical differences in maintenance skills (medium exchange frequency with the combination of manual removal of morphologically irregular colonies). By measuring the all existing colonies from real-time microscopic images, the heterogenous change of colony morphologies in the culture vessel was visualized. By such visualization with morphologically categorized Manhattan chart, the difference between technical skills could be compared for evaluating appropriate cell processing. (C) 2017, The Japanese Society for Regenerative Medicine. Production and hosting by Elsevier B.V.