We propose a microfluidic experimental platform for producing size-controlled spheroids. Cells were aggregated into chambers arranged in an array by microrotational flow within 120 s to form spheroids. The cell density of the initial medium and hydrodynamic flow in the developed array could be adjusted while keeping the device geometry the same to control spheroid size with a standard deviation of less than 19% of the mean. Using this device, spheroids of HepG2 cells of various size categories could be maintained for three days in the chamber with medium exchange and could be continuously evaluated for topology and hepatic functions. Furthermore, CYP1A1 activities were found to increase with time to a constant level at three days. These results demonstrate that this device is readily applicable to producing and maintaining spheroids for in vitro drug screening and biological research.
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