An improved procedure for the isolation of rat brown adipose tissue cells

A method for the isolation of brown adipocytes free of fat interferences and sensitive to noradrenalin is presented. The cells were isolated from pieces of brown adipose tissue with a collagenase treatment. The cells were obtained in the presence of heparin, in order to free the lipoprotein lipase attached to the cell surface. The cells were isolated in the presence of Amberlite XAD-2 [polymeric hydrophobic absorbent beads], which retained most of the fat droplets, formed from broken cells, during the process of disaggregation. The combined use of heparin and Amberlite XAD-2 during the isolation procedure resulted in a lowered cell basal oxygen consumption rate when compared with that of cells isolated with standard methods. The treatment presented lowered the availability of extracellular fatty acids for the isolated brown fat cells, resulting in lower operation of the thermogenin shunt, and thence in decreased basal oxygen consumption and higher sensitivity to glucose and noradrenalin stimulation.