![]() In addition, understanding adipogenesis and the mechanisms that drive adipocyte differentiation requires a cell system that allows detailed temporal assessments and the ability to follow individual cells/groups of cells ( Fig. However, studies of mature adipocytes are hampered by a number of technical hurdles, including high lipid content of these cells that makes them float in suspension, their short life ex vivo span, and the often harsh methods that are required to isolate them, which may impact on their transcriptional and functional fingerprint. Therefore, functional studies of adipocytes and their (dys)regulation under different conditions are of obvious interest for the understanding of systemic metabolic regulation. Induction of BAT thermogenesis as well as transdifferentiation followed by activation of white into brown-like (so-called beige or brite for brown-in-white) adipocytes has raised interest as a therapeutic strategy ( 1). Brown adipose tissue (BAT) is specialized in energy dissipation as heat. Moreover, changes in adipocyte lipid storage (uptake and release) are believed to play a central role in promoting ectopic lipid deposition in tissues like liver and skeletal muscle, which can ultimately lead to lipotoxicity and insulin resistance in these tissues ( 2). Nevertheless, adipocytes constitute almost 90% of the volume and up to 40% of the cells present in WAT ( 1). For example, the role of infiltrating immune cells in attenuating insulin sensitivity is well documented. In addition, WAT displays distinct cell heterogeneity. The close relationship between WAT mass and common cardiometabolic disorders such as insulin resistance, type 2 diabetes, and hypertension has prompted intense research into the mechanisms driving WAT plasticity and phenotype. White adipose tissue (WAT) displays a remarkable capacity for undergoing changes in size and morphology. Capturing adipocyte heterogeneity at the single-cell level within a single fat depot will be key to understanding diversities in cardiometabolic parameters among lean and obese individuals. Further understanding of fat cell physiology and dysfunction will be achieved through genetic manipulation, notably CRISPR-mediated gene editing. Challenges for the future include isolation and culture of adipose-derived stem cells from different anatomic location in animal models and humans differing in sex, age, fat mass, and pathophysiological conditions. An important development is the advent of three-dimensional culture, notably of adipose spheroids that recapitulate in vivo adipocyte function and morphology in fat depots. Here, we survey various models of differentiated preadipocyte cells and primary mature adipocyte survival describing main characteristics, culture conditions, advantages, and limitations. Cultures of mouse and human-differentiated preadipocyte cell lines and primary cells have been established to mimic white, beige, and brown adipocytes. Moreover, a fully differentiated state, notably acquirement of the unilocular lipid droplet of white adipocyte, has so far not been reached in two-dimensional culture. Mature adipocytes float in suspension culture due to high triacylglycerol content and are fragile. The intrinsic properties of adipocytes pose specific challenges in culture. Moreover, adipocytes release lipids and proteins with paracrine and endocrine functions. White, beige, and brown adipocytes differ in their handling of lipids and thermogenic capacity, promoting differences in size and morphology. Several types of fat cells with distinct metabolic properties coexist in various anatomically defined fat depots in mammals. Adipocytes are specialized cells with pleiotropic roles in physiology and pathology.
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