Differentiation and Induction of Lipid Droplet Accumulation in Human Adipocytes as a Model to Study Obesity In Vitro.

Obesity-related changes in adipose tissue alter local secretory profiles and influence interactions with other surrounding cells, including cancer cells. The continued delivery of nutrients shifts the metabolism of fat cells, making them act as a lipid reservoir. Typically, research on obesity mechanisms relies on animal models, which, however, do not allow analysis of metabolic processes at the cellular level. Hence, there is a need to develop a reliable in vitro model of human obesity that can help explore changes in the function of obese adipocytes compared to their lean counterparts. This work presents a protocol for the differentiation of human adipocytes from their precursors (adipose-derived stem cells) using a mixture of insulin, isobutyl-1-methylxanthine, dexamethasone, and indomethacin. Subsequently, differentiated adipocytes were incubated with a set of fatty acids, including linoleate, palmitate, and oleate, to promote lipid accumulation. As a result, a valuable model mimicking the hypertrophic cells under obesity conditions was achieved, which can be used for further studies.