Customized small-sized clinostat using 3D printing and novel polydimethylsiloxane culture dish

Abstract Over the past few decades, research on life in space has increased. Owing to the expensive nature of and challenges associated with conducting experiments in real space, clinostats, which create rotational motion by using motors to reduce the effect of gravity, are used to generate simulated microgravity (SMG) on Earth. However, the existing clinostat systems are complex, large, and expensive, which reduces their accessibility to researchers. Here, by using a 3D printing method, we develop a novel customized small-sized clinostat (CS clinostat) that is easy to manufacture, inexpensive, and robust. The dimensions of the CS clinostat are 282 × 140 × 252 mm3, and therefore, it can fit inside a typical 50 L incubator. The two motors of the CS clinostat rotate at 4 rpm (inner axis) and 1.8 rpm (outer axis) such that the accumulated three-axis acceleration is less than 3 × 10-2 G within 1 h. Moreover, we develop and fabricate a novel culture dish that fits inside the CS clinostat. This dish is covered with polydimethylsiloxane to facilitate gas exchange during cell culture. To validate SMG generation in the CS clinostat, we applied it to mammalian cells, OV-90, TOV-21G, and Caov-3, ovarian cancer cells. Western blotting analysis demonstrated significant reduction in Caveolin-1 expression, a biomarker of SMG, indicating SMG generation. The proposed CS clinostat system has good accessibility and reduces the barriers to SMG research, which makes it useful as a tool for biologists, who are unfamiliar with conventional clinostat equipment, to conduct preliminary studies in the space environment.