Although the effect of molecular hydrogen (H₂) as an antioxidant, etc. on living organisms has garnered considerable attention in recent years. In this study, we developed a culture vessel using a hydrogen gelatin with a high hydrogen gas concentration and investigated the effect of hydrogen on the hepatocellular toxicity of acetaminophen (N-acetyl-p-aminophenol; APAP), because overdosed APAP is known to generate active oxygen in hepatocytes and exhibit hepatotoxicity.

This hydrogen gelatin can contain 36.5 v/w% of H₂, as well as air. The culture vessel consists of a lower container containing hydrogen gelatin and an upper container for cell culture, and these containers are partitioned by a polydimethylsiloxane (PDMS) membrane. Hydrogen was supplied from the pores formed in the PDMS membrane to the upper container at 0.13–0.17 mL/h for 6 h after the culture started.

Under hydrogen gelatin conditions, the number of viable cells, which decreased to 84% owing to the 24 h exposure of 20 mM APAP for cell proliferation under air gelatin conditions, were restored up to 96%. Because the gelatin can contain various gases other than hydrogen, the proposed culture vessel appeared to be effective as an in vitro evaluation system for evaluating the effects of various gas molecules on cells.