Cell pairing and polyethylene glycol (PEG)-mediated cell fusion using two-step centrifugation-assisted single-cell trapping (CAScT)

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 cm(2). 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.