Dynamic single cell culture array
Dynamic single cell culture array
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DOI：
10.1039/b605937f
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发表时间：
2006-01-01
期刊：
影响因子：
6.1
通讯作者：
Lee, Luke P.
中科院分区：
文献类型：
--
作者：
Di Carlo, Dino;Wu, Liz Y.;Lee, Luke P.
重要的是要量化个体细胞群体中的行为分布，以达到对细胞过程的更完整的定量理解。改进的单细胞行为的高通量分析要求单个细胞具有可控的细胞-细胞相互作用的统一条件，包括可扩散元素和接触元素。之前已经使用蛋白质微图案化技术构建了用于静态培养贴壁细胞的统一细胞阵列，但缺乏控制扩散分泌物的能力。在这里，我们提出了一种基于微流控的动态单细胞培养阵列，它既可以对单个贴壁细胞进行阵列培养，也可以在单个细胞的均匀环境下动态控制液体灌流。在我们的设备中，不需要表面修饰，细胞加载在不到30秒内完成。该装置由物理U形流体动力学捕获结构阵列组成，其几何形状偏置为仅捕获单个细胞。HeLa细胞在捕获阵列中的粘附率与在对照玻璃衬底上的粘附率相似。此外，细胞死亡率和分裂率与对照实验相当。观察到大约100个单独的分离细胞在跨度类似于1 mm(2)的视野中生长和附着，24小时后超过85%的细胞保持在主要捕获部位内。此外，在陷阱阵列内灌流培养24小时后，90%以上的细胞贴壁，只有5%的细胞发生了凋亡。我们预计将在单细胞药物毒性分析中使用生理相关的灌流剂量，以及细胞信号通路和系统生物学的研究。
It is important to quantify the distribution of behavior amongst a population of individual cells to reach a more complete quantitative understanding of cellular processes. Improved high-through-put analysis of single cell behavior requires uniform conditions for individual cells with controllable cell - cell interactions, including diffusible and contact elements. Uniform cell arrays for static culture of adherent cells have previously been constructed using protein micropatterning techniques but lack the ability to control diffusible secretions. Here we present a microfluidic-based dynamic single cell culture array that allows both arrayed culture of individual adherent cells and dynamic control of fluid perfusion with uniform environments for individual cells. In our device no surface modification is required and cell loading is done in less than 30 seconds. The device consists of arrays of physical U-shaped hydrodynamic trapping structures with geometries that are biased to trap only single cells. HeLa cells were shown to adhere at a similar rate in the trapping array as on a control glass substrate. Additionally, rates of cell death and division were comparable to the control experiment. Approximately 100 individual isolated cells were observed growing and adhering in a field of view spanning similar to 1 mm(2) with greater than 85% of cells maintained within the primary trapping site after 24 hours. Also, greater than 90% of cells were adherent and only 5% had undergone apoptosis after 24 hours of perfusion culture within the trapping array. We anticipate uses in single cell analysis of drug toxicity with physiologically relevant perfused dosages as well as investigation of cell signaling pathways and systems biology.