Differentiation of human adipocyte precursors in a chemically defined serum-free medium

Stromal-vascular cells from the inguinal fat tissue of human (age range 1.5 month-27 years), were able to undergo adipose conversion when cultured in a medium containing insulin, transferrin and triiodothyronine. Between 10 and 20 per cent of the cells changed their morphology and accumulated lipid droplets within 10 to 15 days. In most cultures, differentiated cells were present in clusters. These clustered cells were shown by indirect immunofluorescence to contain lipoprotein lipase (located in the Golgi region) and by histochemistry to contain glycerol-3-phosphate dehydrogenase. The occurrence of both enzymes was assessed directly by determining enzyme activities and the synthesis of triacylglycerol was demonstrated by incorporation of [U-14C]glucose into lipids. Foreskin fibroblasts did not display any of these phenotypes. The development of a serum-free, chemically defined medium for the differentiation of diploid adipocyte precursors from human should be of interest for the characterization of factors involved in the stimulation or inhibition of the differentiation process.