Brown adipose tissue (BAT) is a potential therapeutic target to manage obesity by increasing energy expenditure. Development of a robust human BAT cell culture model to study the mechanisms of differentiation and thermogenesis is an important step for future therapeutic development. Past attempts have involved primary culture models using tissue extracted from around the cervical spine region in individual patients. However the reproducibility of experiments conducted in primary lines derived from different individuals is poor. Immortalising and expanding individual clonally selected cells based on high UCP1 gene expression is a strategy which overcomes poor reproducibility, but in so doing becomes less representative of primary cells and does not reflect the heterogenous composition of human BAT which is a mix of both brown and beige adipocytes. To address these issues, we developed a primary human brown/beige adipose cell culture model using subacromioclavicular adipose tissue from individuals undergoing shoulder surgery. Fibroblastic pre-adipocytes were isolated from 1-3g of tissue from each patient, expanded in culture, then frozen for storage. Multiple aggregate cell lines were then produced via combining lines from 10-15 individuals. ‘Standard’, previously published, differentiation (DMEM containing 2% fetal calf serum, 1µM rosiglitazone or pioglitazone, 1µM insulin, 500µM isobutylmethylxanthine, 25µM dexamethasone) conditions were employed, then systematically modified to optimise adipogenesis and browning. Evidence of adipogenesis and browning were based on the presence of lipid loading (oil red-O staining), adrenergic responsiveness (lipolysis in response to the β-adrenergic agonist isoproterenol) and UCP-1 protein expression (via western blotting). We have developed a robust human BAT primary cell culture model providing a platform for execution of experimental protocols yielding highly reproducible data. This model has been optimised for the study of human physiological BAT function and its pharmacological manipulation, with a view to informing new anti-obesity strategies.