METHODS AND COMPOSITIONS FOR THE DIFFERENTIATION OF HUMAN PREADIPOCYTES INTO ADIPOCYTES

摘要

The present invention provides methods and compositions for the consistent and quantitative differentiation of human preadipocytes isolated from adipose tissue into adipocytes bearing biochemical, genetic, and physiological characteristics similar to that observed in isolated primary adipocytes. The methods of the invention comprise incubating isolated human preadipocytes, plated at least about 25,000 cells/cm<2>, in a medium containing, glucose, a cyclic AMP inducer such as isobutylmethylxanthine or forskolin, a glucocorticoid or glucocorticoid analogue, insulin or an insulin analogue and a PPAR gamma agonist or a RXR agonist. Also provided are methods for preparing three dimensional biomatrices containing adipocytes differentiated by the methods of the invention. The compositions of the invention include human adipocytes differentiated by the methods of the invention, three-dimensional matrices of adipocytes, and transfected adipocytes. The methods and compositions have use in the drug discovery of compounds having relevance to the disease states of diabetes, obesity, and cardiovascular disease and in the studies of these diseases, and in the grafting of fat tissue.