RESUMEN
Microfluidic-based droplet generation approaches require the design of microfluidic chips and a precise lithography process, which require skilled technicians and a long manufacturing time. Here we developed a centrifugal buoyancy-based emulsification (CBbE) method for producing droplets with high efficiency and minimal fabrication time. Our approach is to fabricate a droplet generation module that can be easily assembled using syringe needles and PCR tubes. With this module and a common centrifuge, high-throughput droplet generation with controllable droplet size could be realized in a few minutes. Experiments showed that the droplet diameter depended mainly on centrifugal speed, and droplets with controllable diameter from 206 to 158 µm could be generated under a centrifugal acceleration range from 14 to 171.9 g. Excellent droplet uniformity was achieved (CV < 3%) when centrifugal acceleration was greater than 108 g. We performed digital PCR tests through the CBbE approach and demonstrated that this cost-effective method not only eliminates the usage of complex microfluidic devices and control systems but also greatly suppresses the loss of materials and cross-contamination. CBbE-enabled droplet generation combines both easiness and robustness, and breaks the technical challenges by using conventional lab equipment and supplies.
RESUMEN
Quickly and easily producing uniform populations of microsphere-based 3D cell cultures using droplet-based templating methods has the potential to enable widespread use of such platforms in drug discovery or cancer research. Here, we advance the design of centrifuge-based droplet generation devices, describe the use of this platform for droplet generation with controlled cell occupancy, and demonstrate weeklong culture duration. Using simple-to-construct devices and easily implemented protocols, the initial concentration of encapsulated cells is adjustable up to hundreds of cells per microsphere. This work demonstrates the first instance of using centrifugal droplet-generating devices to produce large numbers of cell-encapsulating microspheres. Applications of this versatile methodology include the rapid formation of templated 3D cell culture populations suitable for suspension culture or large batch bioreactor studies that require uniform populations.
