Isolation of a Lipocyte-Rich Fraction from Rat Liver Nonparenchymal Cells

  • David A. Otto
  • Richard L. Veech


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.


Parenchymal Cell Alcohol Dehydrogenase Nonparenchymal Cell Nonparenchymal Liver Cell Retinol Acetate 
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  1. Berry, M.N., and Friend, D.S., 1969, High-yield preparation of isolated rat liver parenchymal cells: A biochemical and fine structural study, J. Cell Biol., 43: 506.PubMedCrossRefGoogle Scholar
  2. Blouin, A., Bolender, R.P., and Weibel, E.R., 1977, Distribution of organelles and membranes between hepatocytes and nonhepatocytes in the rat liver parenchyma: A stereological study, J. Cell Biol., 72: 441.PubMedCrossRefGoogle Scholar
  3. Bronfenmajer, S., Schaffner, F., and Popper, H., 1966, Fat-storing cells (lipocytes) in human liver, Arch. Path., 82: 447.PubMedGoogle Scholar
  4. Cornell, N.W., Lund, P., Hems, R., and Krebs, H.A., 1973, Acceleration of gluconeogenesis from lactate by lysine, Biochem. J., 134: 671.Google Scholar
  5. Crow, K.E., Cornell, N.W., and Veech, R.L., 1977, The rate of ethanol metabolism in isolated rat hepatocytes, Alcoholism: Clin. Exp. Res., 1: 43.CrossRefGoogle Scholar
  6. Dileepan, K.N., Wagle, S.R., Hofmann, F., and Decker, K., 1979, Distribution profile of glucokinase and hexokinase in parenchymal and sinusoidal cells or rat liver during development, Life Sci, 24: 89.PubMedCrossRefGoogle Scholar
  7. Kent, G., Gay, S., Inouye, T., Bahu, R., Minick, O.T., and Popper, H., 1976, Vitamin A-containing lipocytes and formation of type III collagen in liver injury, Proc. Natl. Acad. Sci. USA, 73: 3719.PubMedCrossRefGoogle Scholar
  8. Kent, G., Inouye, T., Minick, O.T., and Bahu, R.M., 1977, Role of lipocytes (perisinusoidal cells) in fibrogenesis, in: “Kupffer Cells and Other Liver Sinusoidal Cells,” E. Wisse, and D.L. Knook, eds., p. 73, Elsevier/North-Holland Biomedical Press, Amsterdam.Google Scholar
  9. Knook, D.L., and Sleyster, E.CH., 1976, Separation of kupffer and endothelial cells of the rat liver by centrifugal elutriation, Exp. Cell Res., 99: 444.PubMedCrossRefGoogle Scholar
  10. Knook, D.L., and Sleyster, E.CH., 1977, Preparation and character- ization of kupffer cells from rats and mouse liver, in: “Kupffer Cells and Other Liver Sinusoidal Cells,” E. Wisse and D.L. Knook, eds., p. 273, Elsevier/North-Holland Biomedical Press, Amsterdam.Google Scholar
  11. Krebs, H.A., and Henseleit, K., 1932, Untersuchungen über die harnstoffbildung im tierkörper, Hoppe-Seyler’s Z. Physiol. Chem., 210: 33.CrossRefGoogle Scholar
  12. Krebs, H.A., Cornell, N.W., Lund, P., and Hems, R., 1974, Isolated liver cells as experimental material, in: “Regulation of Hepatic Metabolism,” F. Lundquist and N. Tygstrup, eds., p. 726, Academic Press, New York.Google Scholar
  13. McGee, J.O.’D., and Patrick, R.S., 1972, The role of perisinusoidal cells in hepatic fibrogenesis: An electron microscopic study of acute carbon tetrachloride liver injury, Lab. Invest., 26: 429.Google Scholar
  14. MOrland, J., Bessesen, A., and Svendsen, L., 1979, Incorporation of labelled amino acids into proteins of isolated parenchymal and nonparenchymal rat liver cells in the absence and presence of ethanol, Biochim. Biophys. Acta, 561: 464.PubMedCrossRefGoogle Scholar
  15. Munthe-Kaas, A.C., and Seglen, P.O., 1974, The use of Metrizamide as a gradient medium for isopycnic separation of rat liver cells, FEBS. Lett., 43: 252.PubMedCrossRefGoogle Scholar
  16. Neeld, J.B., and Pearson, W.N., 1963, Macro-and micromethods for the determination of serum vitamin A using trifluoroacetic acid, J. Nutr., 79: 454.PubMedGoogle Scholar
  17. Popper, H., 1944, Distribution of vitamin A in tissue as visualized by fluorescence microscopy, Physiol. Rev., 24: 205.Google Scholar
  18. Popper, H., and Udenfriend, S., 1970, Hepatic fibrosis:correlation of biochemical and morphologic investigation, Am. J. Med., 49: 707.PubMedCrossRefGoogle Scholar
  19. Schnack, H., Stockinger, L., and Wewalka, F., 1967, Adventitous connective tissue cells in the space of Disse and their relation to fibre formation, Rev. Int. Hepatol., 17: 855.PubMedGoogle Scholar
  20. Tanikawa, K., 1975, Ultrastructure of hepatic fibrosis and fat- storing cells, in: “Collagen Metabolism in the Liver,” H. Popper, K. Becker, eds., p. 93, Stratton International Medical Book Corp., New York.Google Scholar
  21. Wake, K., 1971, “Sternzellen” in the liver: Perisinusoidal cells with special reference to storage of vitamin A, Am. J. Anat., 132:429.Google Scholar
  22. Wake, K., 1974, Development of vitamin A-rich droplets in multi-vesicular bodies of rat liver stellate cells, J. Cell Biol., 63: 683.PubMedCrossRefGoogle Scholar
  23. Wisse, E., 1977, Ultrastructure and function of kupffer cells and other sinusoidal cells in the liver, in: “Kupffer Cells and Other Liver Sinusoidal Cells,” E. Wisse and D.L. Knook, eds., p. 33, Elsevier/North-Holland Biomedical Press, Amsterdam.Google Scholar
  24. Zachman, R.D., and Olson, J.A., 1961, A comparison of retinene reductase and alcohol dehydrogenase of rat liver, J. Biol. Chem., 236: 2309.PubMedGoogle Scholar
  25. Zahlten, R.N., Stratman, F.W., and Lardy, H.A., 1973, Regulation of glucose synthesis in hormone-sensitive isolated rat hepatocytes, Proc. Natl. Acad. Sci. USA, 70: 3213.PubMedCrossRefGoogle Scholar
  26. Zahlten, R.N., Hagler, H.K., Nejtek, M.E., and Day, C.J., 1978, Morphological characterization of kupffer and endothelial cells of rat liver isolated by counterflow elutriation, Gastroenterol, 75: 80.Google Scholar

Copyright information

© Springer Science+Business Media New York 1980

Authors and Affiliations

  • David A. Otto
    • 1
  • Richard L. Veech
    • 1
  1. 1.Laboratory of MetabolismNational Institute on Alcohol Abuse and AlcoholismRockvilleUSA

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