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We propose a microfluidic experimental platform for producing size-controlled spheroids. Cells are collected to form a spheroid in chambers by micro-rotational flow in two minutes. The developed array could control the size of three-dimensional spheroids hydrodynamically with the standard deviations less than 19% by varying the cell density of the medium without altering the device geometry. Using the developed perfusion system we experimentally observed that the sizes of formed spheroids remained constant for two days and that the detoxification enzyme, CYP1A1, activities increased.