Protein Fluidics, Burlingame, CA, USA.
Molecular Devices, San Jose, CA, USA.
SLAS Technol. 2021 Jun;26(3):237-248. doi: 10.1177/24726303211000688. Epub 2021 Mar 30.
There is an increasing interest in using three-dimensional (3D) cell structures for modeling tumors, organs, and tissue to accelerate translational research. We describe here a novel automated organoid assay system (the Pu·MA System) combined with microfluidic-based flowchips that can facilitate 3D cell-based assays. The flowchip is composed of sample wells, which contain organoids, connected to additional multiple wells that can hold various assay reagents. Organoids are positioned in a protected chamber in sample wells, and fluids are exchanged from side reservoirs using pressure-driven flow. Media exchange, sample staining, wash steps, and other processes can be performed without disruption to or loss of 3D sample. The bottom of the sample chamber is thin, optically clear plastic compatible with high-content imaging (HCI). The whole system can be kept in an incubator, allowing long-term cellular assays to be performed. We present two examples of use of the system for biological research. In the first example, cytotoxicity effects of anticancer drugs were evaluated on HeLa and HepG2 spheroids using HCI and vascular endothelial growth factor expression. In the second application, the flowchip system was used for the functional evaluation of Ca oscillations in neurospheroids. Neurospheres were incubated with neuroactive compounds, and neuronal activity was assessed using Ca-sensitive dyes and fast kinetic fluorescence imaging. This novel assay system using microfluidics enables automation of 3D cell-based cultures that mimic in vivo conditions, performs multidosing protocols and multiple media exchanges, provides gentle handling of spheroids and organoids, and allows a wide range of assay detection modalities.
人们越来越感兴趣地使用三维（3D）细胞结构来模拟肿瘤、器官和组织，以加速转化研究。我们在这里描述了一种新颖的自动化类器官分析系统（Pu·MA 系统），该系统结合了微流控芯片，可以促进基于 3D 细胞的分析。该流片由含有类器官的样品孔组成，与可以容纳各种分析试剂的其他多个孔相连。类器官位于样品孔的保护室内，使用压力驱动的流动从侧储器交换流体。可以在不中断或损失 3D 样品的情况下进行介质交换、样品染色、洗涤步骤和其他过程。样品室底部是薄的、光学透明的塑料，与高内涵成像（HCI）兼容。整个系统可以放在培养箱中，允许进行长期细胞分析。我们展示了该系统在生物学研究中的两个应用示例。在第一个示例中，使用 HCI 和血管内皮生长因子表达评估了抗癌药物对 HeLa 和 HepG2 球体的细胞毒性作用。在第二个应用中，该流片系统用于神经球体中 Ca 振荡的功能评估。用神经活性化合物孵育神经球体，并使用 Ca 敏感染料和快速动力学荧光成像评估神经元活性。这种使用微流控技术的新型分析系统能够自动化模拟体内条件的 3D 细胞培养，执行多剂量方案和多次介质交换，对球体和类器官进行温和处理，并允许广泛的分析检测模式。
