Simplifying adipogenic differentiation – a step towards a food compatible media

Cultivated meat is a promising technology with the potential to mitigate the ethical and environmental problems associated with traditional meat. Fat plays a key role in meat flavour and texture, however little academic research has focused on fat production resulting in a paucity of adequate adipogenesis protocols for most livestock-derived cells. The development of a suitable protocol is essential to pave the way for cultivated fat as an ingredient, as the traditional cocktail of IBMX, dexamethasone, insulin and rosiglitazone is not food-compatible. Here, we investigate the necessity of these four classic adipogenesis inducers in a serum-containing and an in-house developed serum-free medium (DMAD). Using a full-factorial design, we show that only two of these four inducers, insulin and rosiglitazone, are necessary for adipogenic differentiation, as measured by gene expression changes and lipid accumulation. Additionally, two glucocorticoid receptor binding molecules, progesterone and hydrocortisone, found in DMAD and FBS, are also required. Importantly, this protocol also leads to mature adipocytes in 3D, edible hydrogels. This was demonstrated in both media types and in four species; ruminant and monogastric. We therefore propose this as an alternative protocol for adipogenesis, which, given the replacement of rosiglitazone by a food-compatible PPAR? agonist, can be suitable for human consumption. Overall design: 3 different MSC isolations from Bos Taurus were differentiated in media where each reduced inducer was taken out or when all of them were present (rDM), after 12 days cells were harvested for sequencing.