Skip to main content
Log in

Primary cultures of endothelial cells of the rat liver

A model for ultrastructural and functional studies

  • Published:
Cell and Tissue Research Aims and scope Submit manuscript

    We’re sorry, something doesn't seem to be working properly.

    Please try refreshing the page. If that doesn't work, please contact support so we can address the problem.

Summary

A new isolation and purification procedure for endothelial cells of the rat liver and the conditions for large scale survival of these cells in maintenance culture are reported. Cells isolated by this new method and cultured with homologous rat serum on a collagen matrix show the restoration of several ultrastructural characteristics typical of rat liver endothelial cells in situ, including the broad cytoplasmic extensions that contain the sieve plates. These fenestrated cytoplasmic projections, which cover the liver sinusoids in vivo, are well preserved and are reformed in a manner reminiscent of the situation in situ. Reformation of specific membrane receptors is indicated by the reappearance of the capacity to take up horseradish peroxidase by adsorptive endocytosis, a characteristic that is lost during the cell isolation procedure. From the results obtained in this study, maintenance culture of rat liver endothelial cells seems to be a promising system for studying the regulation of pore size of the fenestrated sieve plates by alcohol and certain hormones, for studying the interaction of endothelial cells with other liver cells and tumor cells, and for studying the mechanisms of adsorptive endocytosis.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Blouin A, Bolender RP, Weibel ER (1977) Distribution of organelles and membranes between hepatocytes and nonhepatocytes in the rat liver parenchyma. J Cell Biol 72:441–455

    Google Scholar 

  • Brouwer A, Knook DL (1977) Quantitative determination of endocytosis and intracellular digestion by rat liver Kupffer cells in vitro. In: Wisse E, Knook DL (eds) Kupffer cells and other liver sinusoidal cells. Elsevier/North-Holland Biomedical Press, Amsterdam, p 343–352

    Google Scholar 

  • Brouwer A, Wanson J-C, Mosselmans R, Knook DL, Drochmans P (1980) Morphology and lysosomal enzyme activity of primary cultures of rat liver sinusoidal cells. Biol Cell 37:35–44

    Google Scholar 

  • Davison PM, Bensch K, Karasek MA (1980) Growth and morphology of rabbit marginal vessel endothelium in cell culture. J Cell Biol 85:187–198

    Google Scholar 

  • Fraser R, Bowler LM, Day WA (1980) Damage of rat liver sinusoidal endothelium by ethanol. Pathology 12:371–376

    Google Scholar 

  • Hofmann F, Decker K (1979) Comparative metabolic studies on liver sinusoidal cells and different types of macrophages. Hoppe-Seyler's Z Physiol Chem 360:905–912

    Google Scholar 

  • Hubbard AL, Wilson G, Ashwell G, Stukenbrok H (1979) An electron microscope autoradiographic study of the carbohydrate recognition systems in rat liver (I). J Cell Biol 83:47–64

    Google Scholar 

  • Knook DL, Sleyster ECh (1976) Separation of Kupffer cells and endothelial cells of the rat liver by centrifugal elutriation. Exp Cell Res 99:444–449

    Google Scholar 

  • Knook DL, Sleyster ECh (1977) Preparation and characterization of Kupffer cells from rat and mouse liver. In: Wisse E, Knook DL (eds) Kupffer cells and other liver sinusoidal cells. Elsevier/North-Holland Biomedical Press, Amsterdam, p 273–288

    Google Scholar 

  • Knook DL, Blansjaar N, Sleyster ECh (1977) Isolation and characterization of Kupffer and endothelial cells from the rat liver. Exp Cell Res 109:317–329

    CAS  PubMed  Google Scholar 

  • Knook DL, Sleyster ECh (1980) Isolated parenchymal, Kupffer and endothelial rat liver cells characterized by their lysosomal enzyme content. Biochem Biophys Res Commun 96:250–257

    Google Scholar 

  • Munthe-Kaas AC (1977) Uptake of macromolecules by rat Kupffer cells in vitro. Exp Cell Res 107:55–62

    Google Scholar 

  • Munthe-Kaas AC, Kaplan G, Seljelid R (1976) On the mechanism of internalization of opsonized particles by rat Kupffer cells in vitro. Exp Cell Res 103:201–212

    Google Scholar 

  • Murakami T (1978) Tannin-osmium conductive staining of biological specimens for non-coated scanning electron microscopy. Scanning 1:127–129

    Google Scholar 

  • Naito M, Wisse E (1978) Filtration effect of endothelial fenestrae on chylomicron transport in neonatal rat liver sinusoids. Cell Tissue Res 190:371–382

    Google Scholar 

  • Praaning-van Dalen DP, Brouwer A, Knook DL (1981) Clearance capacity of rat liver Kupffer, endothelial and parenchymal cells. Gastroenterol 81:1036–1044

    Google Scholar 

  • Rodman JS, Schlesinger P, Stahl P (1978) Rat plasma clearance of horseradish peroxidase and yeast invertase is mediated by specific recognition. FEBS letters 85:345–348

    Google Scholar 

  • Roos E (1981) Cellular interactions during the formation of liver metastases. Thesis, University of Amsterdam

  • Sherer GK, LeRoy EC (1978) Cultivation of microvascular endothelial cells from human preputial skin. J Cell Biol 79:69a

    Google Scholar 

  • Sleyster EC, Westerhuis FG, Knook DL (1977) The purification of nonparenchymal liver cell classes by centrifugal elutriation: In Wisse E, Knook DL (eds) Kupffer cells and other liver sinusoidal cells. Elsevier/North-Holland Biomedical Press, Amsterdam, p 289

    Google Scholar 

  • Steer CJ, Kusiak JW, Brady RO, Jones EA (1979) Selective hepatic uptake of human β-hexosaminidase A by a specific glycoprotein recognition system on sinusoidal cells. Proc Natl Acad Sci 76:2774–2778

    Google Scholar 

  • Steffan A-M, Lecerf F, Keller F, Cinqualbre J, Kirn A (1981) Isolement et culture de cellules endothéliales de foies humain et murin. C R Acad Sc Paris 292:809–815

    Google Scholar 

  • Van Berkel TJC (1979) The role of non-parenchymal cells in liver metabolism. TIBS September: 202–205

  • Van Dierendonck JH, Van der Meulen J, De Zanger RB, Wisse E (1979) The influence of hormones on the diameter of the fenestrae in the endothelial lining of rat liver sinusoids: a SEM study. Ultramicroscopy 4:149

    Google Scholar 

  • Wanson J-C, Mosselmans R, Brouwer A, Knook DL (1979) Interaction of adult rat hepatocytes and sinusoidal cells in coculture. Biol Cell 36:7–16

    Google Scholar 

  • Wisse E (1970) An electron microscopic study of the fenestrated endothelial lining of rat liver sinusoids. J Ultrastruct Res 31:125–150

    Google Scholar 

  • Wisse E (1972) An ultrastructural characterization of the endothelial cell in the rat liver sinusoid under normal and various experimental conditions, as a contribution to the distinction between endothelial and Kupffer cells. J Ultrastruct Res 38:528–562

    Google Scholar 

  • Wisse E (1974) Observations on the fine structure and peroxidase cytochemistry of normal rat liver Kupffer cells. J Ultrastruct Res 46:393–426

    Google Scholar 

  • Wisse E (1977) Ultrastructure and function of Kupffer cells and other sinusoidal cells in the liver. In: Wisse E, Knook DL (eds) Kupffer cells and other liver sinusoidal cells. Elsevier/North-Holland Biomedical Press, Amsterdam, p 33

    Google Scholar 

  • Wisse E, Knook DL (1979) The investigation of sinusoidal cells: A new approach to the study of liver function. In: Popper H, Schaffner F (eds) Progress in liver diseases Vol VI. Grune and Stratton, New York, p 153–171

    Google Scholar 

  • Wisse E, Roels F, De Prest D, Van der Meulen J, Emeis JJ, Daems WT (1974) Peroxidatic reaction of Kupffer and parenchymal cells compared in rat liver. In: Wisse E, Daems WT, Molenaar I, Van Duijn P (eds) Electron microscopy and cytochemistry. North-Holland, Amsterdam, p 119–122

    Google Scholar 

  • Wisse E, Gregoriadis G, Daems WTh (1976) Electron microscopic cytochemical localization of intravenously injected liposome-encapsulated horseradish peroxidase in rat liver cells. In: Sherwood MR, Escobar MR, Friedman H (eds) The reticuloendothelial system in health and disease. Functions and characteristics. Plenum Publishing Corporation, New York, p 237–245

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

de Leeuw, A.M., Barelds, R.J., de Zanger, R. et al. Primary cultures of endothelial cells of the rat liver. Cell Tissue Res. 223, 201–215 (1982). https://doi.org/10.1007/BF00221510

Download citation

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00221510

Key words

Navigation