Isolation, Culturing and in Vitro Activation of Liver Macrophages

  • Gerrit L. Scherphof
  • Toos Daemen
  • Johannes T. P. Derksen
Part of the NATO ASI Series book series (NSSA, volume 218)


Systemic administration of particulate drug carriers such as liposomes generally leads to rapid accumulation of the carrier in cells of the mononuclear phagocyte system (MPS) (Scherphof et al., 1983). The mechanism of entry is most likely to be of endocytic nature (Scherphof et al., 1985), implying that the carrier-drug complex will be subject to intralysosomal processing. For a drug to be released from the lysosomal compartment in an active or activatable form it will have to be dissociated from the carrier and pass the lysosomal membrane when, as will most often be the case, its action is required in the cytosol. The great majority of the MPS is concentrated in the liver and the spleen, the organs which usually are responsible for the bulk of the removal of the drug carrier from the circulation. In order to be able to study the intracellular fate of such drug carriers as a function of various carrier parameters such as size and composition without being bothered by the involvement, either directly or indirectly, of other cell types, it might be advantageous to perform such studies in vitro with isolated cells. Since the liver macrophage population comprises the largest population of MPS cells in any one organ and as such contributes most extensively to the elimination of particulate drug carriers from the blood, we set out to investigate in detail the uptake and intracellular processing of one of the most versatile drug carriers, the liposome, by isolated liver macrophages (Kupffer cells) in monolayer culture. Also the action of a biologically active compound, the immunomodulator muramyl dipeptide (MDP), encapsulated in liposomes, was studied in this system.


Kupffer Cell Mononuclear Phagocyte System Muramyl Dipeptide Liver Macrophage Multilamellar Liposome 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Daemen, T., Veninga, A., Roerdink, F.H. and Scherphof, G.L. (1986) In vitro activation of rat liver macrophages to tumoricidal activity by free or liposomeencapsulated muramyl dipeptide. Cancer Res. 46, 4330–4335.PubMedGoogle Scholar
  2. Daemen, T., Veninga, A., Dijkstra, J. and Scherphof, G. (1989a) Differential effects of liposome incorporation on macrophage-activating potencies of LPS, Lipid A and MDP; differences in susceptibility to lysosomal enzymes. J. Immunol. 142, 2469–2474.PubMedGoogle Scholar
  3. Daemen, T., Veninga, A., Roerdink, F.H. and Scherphof, G.L. (1989b) Conditions controlling tumor cytotoxicity of rat liver macrophages mediated by liposomal muramyl dipeptide. Biochim. Biophys. Acta 991, 145–151.PubMedCrossRefGoogle Scholar
  4. Daemen, T., Koudstaal, J., Scherphof, G.L. and Hardonk, M.J. (1989c) Population kinetics of rat liver macrophages after intravenous administration of liposomeencapsulated MDP, in: Cells of the Hepatic Sinusoid, Vol. 2 (E. Wisse, D.L. Knook and K. Decker, eds.) Kupffer Cell Foundation, Rijswijk, The Netherlands, pp. 400–405.Google Scholar
  5. Daemen, T., Veninga, A., Roerdink, F.H. and Scherphof, G.L. (1989d) Endocytic and tumoricidal heterogeneity of rat liver macrophage populations; implications for drug targeting. Select. Cancer Therap., in press.Google Scholar
  6. Derksen, J.T.P. and Scherphof, G.L. (1985) An improved method for the covalent coupling of proteins to liposomes. Biochim. Biophys. Acta 814, 151–155.CrossRefGoogle Scholar
  7. Derksen, J.T.P., Morselt, H.W.M. and Scherphof, G.L. (1987) Processing of different liposome markers after in vitro uptake of immunoglobulin-coated liposomes by rat liver macrophages. Biochim. Biophys. Acta 931, 33–40.PubMedCrossRefGoogle Scholar
  8. Derksen, J.T.P., Morselt, H.W.M. and Scherphof, G.L. (1988) Uptake and processing of immunoglobulin-coated liposomes by subpopulations of rat liver macrophages. Biochim. Biophys. Acta 971, 127–136.PubMedCrossRefGoogle Scholar
  9. Dijkstra, J., Van Galen, W.J.M., Hulstaert, C.E., Kalicharan, D., Roerdink, F.H. and Scherphof, G.L. (1984a) Interaction of liposomes with Kupffer cells in vitro. Exp. Cell Res. 150, 161–176.PubMedCrossRefGoogle Scholar
  10. Dijkstra, J., Van Galen, M. and Scherphof, G. (1984b) Effects of ammoniumchloride and chloroquine on endocytic uptake of liposomes by Kupffer cells in vitro. Biochim. Biophys. Acta 804, 58–67.PubMedCrossRefGoogle Scholar
  11. Dijkstra, J., Van Galen, M., Regts, J. and Scherphof, G. (1985a) Uptake and processing of liposomal phospholipids by Kupffer cells in vitro. Eur. J. Biochem. 148, 391–397.PubMedCrossRefGoogle Scholar
  12. Dijkstra, J., Van Galen, M. and Scherphof, G. (1985b) Effects of (dihydro)cytochalasin B, colchicin, monensin and trifluoperazine on uptake and processing of liposomes by Kupffer cells in culture. Biochim. Biophys. Acta 845, 34–42.PubMedCrossRefGoogle Scholar
  13. Fidler, I.J., Sone, S., Fogler, W.E. and Barnes, Z.L. (1981) Eradication of spontaneous metastases and activation of alveolar macrophages by intravenous injection of liposomes containing muramyl dipeptide. Proc. Natl. Acad. Sci. USA 78, 1680–1684.PubMedCrossRefGoogle Scholar
  14. Fidler, I.J., (1980) Therapy of spontaneous metastases by intravenous injection of liposomes containing lymphokines. Science 208, 1469–1471.PubMedCrossRefGoogle Scholar
  15. 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, 444–449.PubMedCrossRefGoogle Scholar
  16. Olson, F., Hunt, C.A., Szoka, F.C., Vail, W.J. and Papahadjopoulos, D. (1979) Preparation of liposomes of defined size distribution by extrusion through polycarbonate membranes. Biochim. Biophys. Acta 557, 9–23.PubMedCrossRefGoogle Scholar
  17. Roerdink, F., Regts, J. and Scherphof, G. (1986) Effect of lipid composition on the uptake and intracellular degradation of liposomes by Kupffer cells, in: Cells of the hepatic sinusoid (A. Kirn, D.L. Knook and E. Wisse, eds.) Kupffer Cell Foundation, Rijswijk, pp. 131–136.Google Scholar
  18. Roerdink, F.H., Regts, J., Handel, T., Sullivan, S.M., Baldeschwieler, J.D. and Scherphof, G.L. (1989) Effect of cholesterol on the uptake and intracellular degradation of liposomes by liver and spleen; a combined biochemical and gammaray perturbed angular correlation study. Biochim. Biophys. Acta 980, 234–240.PubMedCrossRefGoogle Scholar
  19. Scherphof, G., Roerdink, F., Dijkstra, J., Ellens, H., De Zanger, R. and Wisse, E. (1983) Uptake of liposomes by rat and mouse hepatocytes and Kupffer cells. Biol. Cell 47, 47–58.Google Scholar
  20. Scherphof, G.L., Dijkstra, J., Spanjer, H.H., Derksen, J.T.P. and Roerdink, F.H. (1985) Uptake and intracellular processing of targeted and non-targeted liposomes by rat Kupffer cells in vivo and in vitro. Ann. N.Y. Acad. Sci. 446, 368–384.PubMedCrossRefGoogle Scholar
  21. Stewart, C.C., Stevenson, A.P. and Hibbs, J. (1988) Effector mechanisms for macrophage-induced cytostasis and cytolysis of tumor cells, in: Macrophages and Cancer (G. Heppner and A.M. Fulton, eds.) CRC Press, pp. 40-59.Google Scholar

Copyright information

© Plenum Press, New York 1991

Authors and Affiliations

  • Gerrit L. Scherphof
    • 1
  • Toos Daemen
    • 1
  • Johannes T. P. Derksen
    • 1
  1. 1.Laboratory of Physiological ChemistryState University GroningenGroningenThe Netherlands

Personalised recommendations