Here a convenient and effective strategy of two-step centrifugation-assisted single-cell trapping (CAScT) based on an addressable truncated cone-shaped microwell array (TCMA) chip is developed for cell pairing. We describe the operation principles of the method and demonstrate its compatibility with polyethylene glycol (PEG)-mediated cell fusion. Compared with other methods, most of which rely on sophisticated devices and bulky subsidiary equipment, our method is more convenient and exhibits better or comparable performance. Using this method, up to around 6000 heterotypic cell pairs can be formed and addressed within a small area of 1 cm². The paired cells are then treated with fusogenic PEG for cell fusion. Compared with traditional protocols, cell fusion using this approach is well defined with better control, which leads to an improved yield of heterotypic binucleated hybrids. Furthermore, we demonstrate the capability of our device for long-term cell culture and cell harvesting.

中文翻译：

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使用两步离心辅助单细胞捕获（CAScT）进行细胞配对和聚乙二醇（PEG）介导的细胞融合†

在这里，一种方便有效的策略是基于可寻址的截短圆锥形微孔阵列（TCMA）芯片的两步离心辅助单细胞捕获（CAScT），用于细胞配对。我们描述了该方法的操作原理，并证明了其与聚乙二醇（PEG）介导的细胞融合的相容性。与其他方法（大多数方法都依赖于复杂的设备和笨重的辅助设备）相比，我们的方法更方便，并且表现出更好的性能或可比的性能。使用这种方法，可以在1 cm ²的小区域内形成并寻址多达约6000个异型细胞对。。然后用融合PEG处理配对的细胞以进行细胞融合。与传统方案相比，使用这种方法进行的细胞融合得到了更好的控制，并得到了更好的定义，从而提高了异型双核杂种的产量。此外，我们展示了我们的设备用于长期细胞培养和细胞收获的能力。