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Surface area ratios. III. A stereological method for estimating relative changes in average hepatocytes

Stereological information seems to be most interpretable, with respect to experimental changes, when related to an average cell. Current methods for obtaining an average cell volume essentially consist of dividing an aggregate volume of cells by the number of nuclei therein, assuming one nucleus per cell (Loud, ′68). Hepatocytes represent a somewhat special case, however, in that some are binucleated. Since the number of hepatocytes in one cm3 of tissue is less than the number of hepatocyte nuclei in the same cm3, dividing the hepatocytic volume by the number of nuclei gives only an average “mononuclear” hepatocyte. Such an estimate creates two interpretation difficulties: (1) the volume of an average mononuclear hepatocyte is less than that of an average hepatocyte; and (2) changes in the proportion of binucleated cells may compromise the “relative comparisons” for which the method was originally intended. The purpose of this study is to describe a new approach that can avoid these difficulties altogether, and then to assess the errors associated with the average mononuclear hepatocyte estimates. This was accomplished by combining a surface area ratio method, which can detect average cell changes without being influenced by binucleated cells, with the method of Loud (′68), which is affected as described above. The experimental model was the rat liver (n = 20) recovering for 3 days from 5 daily injections of sodium phenobarbital (100 mg/kg). The results indicate that changes in the average cell volumes for the two methods have similar slopes, but by not accounting for binucleated cells, the average mononuclear hepatocyte reference overestimates average hepatocytic volume changes by 63.1%. Similarly, the mononuclear hepatocyte reference overestimates changes in the surface areas of the ER by 32.1% (range = 26.1% to 39.1%), the SER by 21.6% (range = 14.3 to 30.1%), and the RER by 65.1% (range = 54.6% to 76.4%).