An extremely simple, self-standing microfluidic cell culture system is reported. The whole system is confined in a 35 mm culture dish, and requires only a standard CO2 incubator. The culture medium is perfused by gravity. We successfully cultured NIH3T3-derived cells up to 10 days with a viability of similar to 90%. (c) 2014, The Society for Biotechnology, Japan. All rights reserved.