Masculinized and feminized physiological cell culture media

Cell culture is an important tool in molecular and cellular physiology. Recently, culture media (e.g. HPLM, Plasmax) modeled after the human blood plasma metabolome have been developed. However, these media are agnostic to biological sex. Generally, sex is often ignored as a variable in cell biology. To effectively include biological sex as a variable in cell culture, media could be developed to reproduce male-like or female-like blood plasma. To address this, we gathered data on the concentrations of 86 plasma metabolites, measured in healthy human adults of both sexes. A meta-analysis of the data indicated that mean concentrations of many amino acids, and a number of other metabolically significant metabolites, are significantly different between males and females. Similarly, a range of steroid hormones also differend significantly. We formulated a ‘masculinized’ and a ‘feminized’ cell culture media based on these values and use these media to culture Molt-4 and Loucy cell lines, which are male and female T-lymphoblast lines, respectively. These experiments are ongoing and will include RNAseq analyses to identify gene expression differences, as well as metabolic assays. Nonetheless, we already have sufficient insight into the male and female plasma metabolomes to understand that, as we move forward with more physiological cell culture approaches, the impact of biological sex must be considered. Natural Science and Engineering Research Council of Canada This abstract was presented at the American Physiology Summit 2025 and is only available in HTML format. There is no downloadable file or PDF version. The Physiology editorial board was not involved in the peer review process.