Isolation of a lipocyte-rich fraction from rat liver nonparenchymal cells.
A modified pronase digestion procedure is described for isolating nonparenchymal liver cells from vitamin A treated rats, which yielded a 15-23% population of lipocytes in the total cell suspension. The criteria for defining a lipocyte was the appearance of vitamin A containing cells as determined by fluorescent microscopy. Measurement of alcohol dehydrogenase and retinol dehydrogenase activities indicated that these enzyme activities were not present in the isolated nonparenchymal cells. A lipocyte-rich fraction of nonparenchymal cells was obtained by centrifugation of purified nonparenchymal cells in a linear Metrizamide gradient. Vitamin A fluorescence and chemical assay of vitamin A in the cell fractions indicated a four-fold enrichment of lipocytes in the cell fraction with d = 1.043 g/ml. Fractions high in vitamin A also had numerous cells with fat droplets as shown by transmission electron microscopy.
D000273:Adipose Tissue / Q000166:cytology*; D000429:Alcohol Oxidoreductases / Q000378:metabolism; D000818:Animals; D002469:Cell Separation / Q000379:methods; D002499:Centrifugation, Density Gradient; D000072467:Cytochrome P450 Family 2; D066298:In Vitro Techniques; D008099:Liver / Q000166:cytology* / Q000201:enzymology; D008297:Male; D051381:Rats; D014801:Vitamin A / Q000378:metabolism
D014801:Vitamin A; D000429:Alcohol Oxidoreductases; C022520:retinol dehydrogenase; C504568:Cyp2d1 protein, rat; D000072467:Cytochrome P450 Family 2