Differentiation Kinetics of in Vitro 3T3-L1 Preadipocyte Cultures

Abstract

Engineering autologous adipose constructs from cell culture is a promising strategy to overcome limitations of conventional soft-tissue implants. A methodology is presented to experimentally determine and mathematically model the differentiation kinetics of in vitro 3T3-L1 preadipocyte cultures that can aid in construct design. Relative rates of morphological and interfacial events during adipogenesis were compared. Model results suggest that maturation of an intermediate multilocular phenotype was the rate-limiting step in morphological differentiation and had an intrinsic rate of 0.012 day^-1. Dislodgment of multilocular fat cells was the primary mechanism of cell loss during adipogenesis. The maximum rate of lipid droplet nucleation was predicted to precede that of coalescence by 10 days and to be three times faster. Coalescence probability was estimated to decrease from 33 to 11% for 4- and 8-μm-diameter droplets, respectively. Fluid drainage and the cytoskeleton between droplets could have impeded coalescence. The kinetic analysis suggests that droplet ripening was the dominant mechanism of lipid production. Applications of this research include engineering of an adipose construct and predicting surgical outcome of patients requiring soft-tissue augmentation.