Establishment of a physiological culture system to improve translation in metabolic research

The major output from this will be technological. The ability to control the oxidative status of cells, combined with a more physiological cell culture media, will provide a powerful new tool for the study of adipose tissue biology. Furthermore, a deeper understanding of how cell lines respond to their environment has broad implications for other cellular models. This project will set standards for adipocyte models, improving reproducibility and translation between cells, mice and humans.
Improved cellular models for studying metabolic diseases will benefit academic and industrial stakeholders. The fact our proposed technology is cheap and straightforward to implement makes it accessible to a global audience, not just those based in well-funded research environments. We will disseminate information through publication and also the Cambridge Metabolic Network Website to provide more detailed protocols. Research data management (RDM) will follow Findable, Accessible, Interoperable, and Re-usable (FAIR) principles.

To optimise cell culture conditions for modelling adipocyte biology in vitro.
Mammalian cell culture is a staple of laboratories. However, the translatability of cell models to in vivo settings is debated. Research from cancer models suggests improved media formulations can significantly improve translation to in vivo settings1. In the metabolism field, cell lines are widely used to model organs such as muscle, liver and adipose tissue. However, the impact of culture conditions on their phenotypes is underexplored, making optimised cellular models a critical unmet need for metabolic research.
Metabolic cell types are characterised by high oxygen consumption. Despite the prevalent view that cell culture is a state of high oxygen tension2, the diffusion gradient from the media surface to the cell monolayer results in cells such as 3t3-L1 adipocytes and primary hepatocytes being hypoxic3 and highly glycolytic. Furthermore, most culture media are deficient in specific essential vitamins4. We identified that combining increased oxygen tension (by altering media depth) with vitamins (B7/B12) reprograms the 3t3-L1 adipocyte cell line to better model in vivo adipocytes. Notably, hypoxia and vitamin deficiency are both characteristics of obesity and metabolic diseases.
Here we propose to validate our “physiological” 3t3-L1 cultures by deep-phenotyping them using -omics approaches and also extend work to other models (human IPSC-derived adipocytes, brown adipocytes). Comparing these to data from tissue banks of murine and human adipose tissue will allow us to further optimise our in vitro models and maximise translation to in vivo models.