Adipocytes are functional cells specialised in the storing and control of energy stores. While the majority of adipocytes belongs to the unilocular white lineage responsible for lipid storage, a subset of adipogenic cells known as brown adipocytes are able to dissipate energy through thermogenesis. This thermogenic capacity relies on the brown-specific expression of the mitochondrial uncoupling protein UCP1. In vivo, cold exposure is the main stimulus to activate UCP1, known for mediating heat production in brown and beige adipocytes, however the physiological factors favouring thermogenic brown adipogenic lineages in vitro remain largely elusive. In order to identify key biological stimuli involved in adipogenic browning, we have developed an in vitro culture system to induce an adipogenic response using marrow-derived stem cells (MSCs) exposed to defined culture conditions. Under specific treatment, including exposure to reduced temperature during differentiation, MSCs can form adipocytes sharing metabolic, protein and molecular features of brown adipocytes. Incubation at 32°C instead of 37°C over the course of the culture treatment was seen to promote brown adipogenesis, with increased adipocyte response and upregulation of adipogenic and thermogenic factors including UCP1. Treated cultures generated multilocular adipocytes exhibiting a progressive increase in the number and size of intracellular lipid droplets within a week. Ultrastructural analysis indicated that alongside increased intracellular lipid inclusions, treated cells showed number of mitochondria with altered morphology. Quantitative metabolic measurements confirmed that organelle adaptation was accompanied by enhanced basal respiration, increased glycolysis and oxidative phosphorylation, indicating metabolically active cells consistent with a thermogenic phenotype. These latest results demonstrate the usefulness of stem cell cultures available from accessible tissue for the study of adipocyte formation, providing a unique model to measure cellular and metabolic adaptation to pharmacological and environmental stimuli. This in vitro model represents a new tool to identify pro-browning factors at the cellular level, and elucidate the mechanism of metabolic adaptation relevant to the browning process.