PREPARATION OF FAT CELL-"FREE" RAT MAMMARY GLAND

The uptake and clearance of chemical carcinogens by mammary tissue are for the most part obscured by the large amount of fat surrounding the mammary parenchymal tissue. This is due primarily to the lipophilic nature of the carcinogenic polycyclic hydrocarbons. Therefore, it became necessary to develop a procedure whereby mammary gland adipose tissue cells may be separated from mammary parenchymal tissue without removal of intracellular lipid or the disruption of the tissue beyond histologic recognition. The procedure developed involves the incubation of the tissue with the enzyme collagenase in a tissue culture medium for 1-1 ½ hr. This is followed by several sodium chloride washes which effectively remove any adhering intact fat cells. Lipid content of the collagenasetreated preparation was decreased from approximately 60% to 2% when compared with the untreated control gland. The procedure described effectively separates intact fat cells from mammary parenchymal tissue but maintains a histologic state in which mammary ducts and alveoli may be readily identified.