In situ monitoring of 3D in vitro cell aggregation using an optical imaging system
In situ monitoring of 3D in vitro cell aggregation using an optical imaging system
复制标题
使用光学成像系统原位监测 3D 体外细胞聚集
DOI：
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发表时间：
2008
影响因子：
3.8
通讯作者：
S. Morgan
中科院分区：
文献类型：
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作者：
N. Sawyer;L. K. Worrall;J. Crowe;S. L. Waters;K. Shakesheff;F. Rose;S. Morgan
将细胞和组织保持在悬浮液中的生物反应器系统越来越受欢迎地用于培养3D构建体，以避免与传统2D培养方法相关的体内细胞功能的丧失。有必要对这些系统进行在线监测，以便更好地了解和控制所涉及的过程，并防止离线取样和终点分析造成这些过程的中断。我们描述了一个系统的成像和细胞聚集的分析，在很长一段时间内，在一个高方面旋转血管（HARV）。该系统利用侧面照明，使用可调节的光束图案，以限制检测到的光被细胞聚集体散射，从而消除了对细胞的荧光标记的需要。在8小时内监测哺乳动物细胞（MCF-7乳腺癌细胞）的原位聚集，并获得显示生物反应器内聚集体的生长和运动的图像序列。详细的大小和人口数据已经得出的特点，在这段时间内的聚集体的发展。我们展示了可分辨聚集体的数量如何增加到达到峰值，然后随着这些聚集体的合并而下降。一旦形成，剩余的聚集体就会合并形成更紧密堆积的主体，通常横截面积会减少三分之一。这些结果为自动反馈系统的开发提供了基础，以控制3D细胞培养物的生长，从而进行可重复、可靠和质量受控的实验。Biotechnol. Bioeng. 2008;100：159-167.© 2007 Wiley Periodicals，Inc.
Bioreactor systems that maintain cells and tissues in suspension are increasingly popular for culturing 3D constructs to avoid the loss of in vivo cell function associated with traditional 2D culture methods. There is a need for the online monitoring of such systems to provide better understanding and control of the processes involved and to prevent the disruption of these processes caused by offline sampling and endpoint analysis. We describe a system for the imaging and analysis of cell aggregation, over long periods, within a high aspect rotating vessel (HARV). The system exploits side illumination, using an adjustable beam pattern, to restrict the detected light to that scattered by the cell aggregates, thus eliminating the need for the fluorescent labeling of the cells. The in situ aggregation of mammalian cells (MCF‐7 breast carcinoma cells) was monitored over an 8 h period and image sequences showing the growth and motion of the aggregates within the bioreactor were obtained. Detailed size and population data have been derived characterizing the development of the aggregates during this time. We show how the number of resolvable aggregates increases to reach a peak and then declines as these aggregates merge. Once formed, remaining aggregates are found to consolidate to form more tightly packed bodies, typically reducing in cross‐sectional area by one third. These results provide the basis for the development of an automated feedback system to control the growth of 3D cell cultures for repeatable, reliable, and quality controlled experimentation. Biotechnol. Bioeng. 2008;100: 159–167. © 2007 Wiley Periodicals, Inc.