Development of an optical system for the non-invasive tracking of stem cell growth on microcarriers.
Development of an optical system for the non-invasive tracking of stem cell growth on microcarriers.
复制标题
开发用于微载体干细胞生长的非侵入性跟踪的光学系统。
DOI：
10.1002/bit.26328
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发表时间：
2017-09
影响因子：
3.8
通讯作者：
Coopman K
中科院分区：
文献类型：
--
作者：
Odeleye AOO;Castillo-Avila S;Boon M;Martin H;Coopman K
干细胞疗法的医学适应症的出现已经看到了开发贴壁细胞如间充质干细胞（MSC）的制造能力的需要。一个这样的发展是在微载体的使用中，当与2D培养平台相比时，微载体促进贴壁干细胞培养物的细胞密度增加。考虑到商业上可用的干细胞扩增系统的多样性，在不同环境中对细胞数量、汇合和聚集进行非侵入性和自动化监测的新方法将成为过程控制的必要条件，以确保可靠和一致的性能。使用定制显微镜实现了附着于Cytodex 1和3微载体的小鼠胚胎成纤维细胞和人间充质干细胞的原位落射照明。开发了稳健的图像处理技术，以提供汇合、聚集体识别和细胞数量的定量测量，而不需要荧光标记或细胞分离。对单个微载体上计数的细胞的大数据集进行了统计分析，并与NucleoCounter测量结果进行了比较，在聚集开始前，从12天培养的第0天到第6天观察到的平均差异小于7%。所开发的图像采集系统和后处理方法成功应用于动态移动的定殖微载体。所提出的系统提供了一种在个体水平上进行细胞鉴定的新方法，以一致且准确地评估活细胞数量、汇合和细胞分布，同时还最小化了当前侵入性手段中固有的变异性，通过该侵入性手段分析粘附于微载体的细胞。Biotechnol. Bioeng. 2017;114：2032-2042.© 2017作者。生物技术和生物工程出版的威利期刊，公司。
The emergence of medicinal indications for stem cell therapies has seen a need to develop the manufacturing capacity for adherent cells such as mesenchymal stem cells (MSCs). One such development is in the use of microcarriers, which facilitate enhanced cell densities for adherent stem cell cultures when compared with 2D culture platforms. Given the variety of stem cell expansion systems commercially available, novel methods of non‐invasive and automated monitoring of cell number, confluence, and aggregation, within disparate environments, will become imperative to process control, ensuring reliable and consistent performance. The in situ epi‐illumination of mouse embryonic fibroblasts and human mesenchymal stem cells attached to Cytodex 1 and 3 microcarriers was achieved using a bespoke microscope. Robust image processing techniques were developed to provide quantitative measurements of confluence, aggregate recognition, and cell number, without the need for fluorescent labeling or cell detachment. Large datasets of cells counted on individual microcarriers were statistically analyzed and compared with NucleoCounter measurements, with an average difference of less than 7% observed from days 0 to 6 of a 12‐day culture noted, prior to the onset of aggregation. The developed image acquisition system and post‐processing methodologies were successfully applied to dynamically moving colonized microcarriers. The proposed system offers a novel method of cell identification at the individual level, to consistently and accurately assess viable cell number, confluence, and cell distribution, while also minimizing the variability inherent in the current invasive means by which cells adhered to microcarriers are analyzed. Biotechnol. Bioeng. 2017;114: 2032–2042. © 2017 The Authors. Biotechnology and Bioengineering Published by Wiley Periodicals, Inc.
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