Unravelling bovine preadipocyte differentiation and their three-dimensional cultivation for cellular agriculture.

Cultured meat is an attractive concept to address environmental and ethical issues of livestock farming. Replication of conventional meat requires fat tissue, for flavour, texture, and mouthfeel. However, cultured meat production is limited by the lack of bovine adipogenic cells that are both proliferative and differentiable. Here, we demonstrate the isolation and characterisation of spontaneously immortalised bovine preadipocytes proliferating over 100 days and 21 passages, showing adipogenic differentiation. Upon induction, the cells accumulate lipids and express key adipogenic markers. Comparative RNA sequencing and proteomic analysis between induced and non-induced cells confirmed bovine origin, cell identity, and low tumour suppressor expression. Proteomic profiling identified over 2000 proteins, with enrichment in adipogenesis-associated pathways, including PPAR signalling and the ERK1/2 cascade. Additionally, the cells demonstrate viability in suspension and strong scaffold attachment. These results establish a reproducible bovine cell source for cultured fat supporting scale-up and bioprocess research in cellular agriculture.