Summary
Liver connective tissue cells have been characterized as perisinusoidal myofibroblasts and hepatic lipocytes (Ito cells, fat-storing cells). A concept of a single mesenchymal cell population that may be modulated between these two phenotypes has been postulated. We have previously established a continuous murine cell line, GRX, obtained from fibrotic granulomatous lesions induced by schistosomal infection in mouse liver. This cell line is considered to represent liver myofibroblasts. In the present study we have induced the conversion of these cells into lipocyte (fat storing) phenotype by treatment with insulin and indomethacin. We have quantified the lipid synthesis and the increase of activity of involved enzymes during the induction of the fat-storing phenotype and described modifications of cell organization along this modulation of cell functions.
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Ailhaud, G. Adipose cell differentiation in culture. Mol. Cell. Biochem. 49:17–31; 1982.
Alfonzo, M.; Apitz-Castro, R. A new catalytic activity ofa-glycerolphosphate dehydrogenase: the enzymatic hydrolysis ofp-nitrophenyl acetate. FEBS Lett. 19:235–238; 1971.
Bhatal, P. S. Presence of modified fibroblasts in cirrhotic livers in man. Pathology 4:139–144; 1972.
Bioulac-Sage, P.; Balabaud, C. La cellule perisinusoidale (ou cellule de Ito). Gastroenterol. Clin. Biol. 9:312–322; 1985.
Borojevic, R.; Vinhas, S. A.; Monteiro, A. N. A., et al. Liver connective tissue cells isolated from human schistosomal fibrosis or alcoholic cirrhosis represent a modified phenotype of smooth muscle cells. Biol. Cell 53:231–238; 1985.
Borojevic, R.; Monteiro, A. N. A.; Vinhas, S. A., et al. Establishment of a continuous cell line from fibrotic schistosomal granulomas in mice livers. In Vitro Cell. Dev. Biol. 21:382–390; 1985.
Burton, K. Determination of DNA concentration with diphenylamine. In: Grossman, L.; Moldave, K., eds. Methods in enzymology XII New York: Academic Press; 1968:163–165.
Clement, B.; Emonard, H.; Rissel, M., et al. Cellular origin of collagen and fibronectin in liver. Cell. Mol. Biol. 30:489–496; 1984.
Cook, K. S.; Hunt, C. R.; Spiegelman, B. M. Developmentally regulated mRNAs in 3T3-adipocytes: analysis of transcriptional controls. J. Cell Biol. 100:514–520; 1985.
Davis, B. H.; Pratt, B. M.; Madri, J. A. Retinol and extracellular collagen matrices modulate hepatic Ito cells collagen phenotype and cellular retinol-binding protein levels. J. Biol. Chem. 261:10280–10286; 1987.
DeLeeuw, A. M.; McCarthy S. P.; Geerts, A., et al. Purified rat liver fat-storing cells in culture divide and contain collagen. Hepatology 4:392–403; 1984.
Forest, C.; Czerucka, D.; Grimaldi, P., et al. Metabolic characteristics of murine adipocyte precursor clones. In: Angel, A.; Hollenberg, C. H.; Roncari, D. A. K., eds. The adipocyte and obesity: cellular and molecular mechanisms. New York: Raven Press; 1983:53–64.
Franke, W. W.; Hergt, M.; Grund, C. Rearrangement of vimentin cytoskeleton during adipose conversion: formation of an intermediate filament cage around lipid globules. Cell 49:131–141; 1987.
Friedman, S. C.; Roll, F. J.; Boyles, J., et al. Hepatic lipocytes: the principal collagen-producing cells of normal rat liver. Proc. Natl. Acad. Sci. USA 82:8681–8685; 1985.
Green, H.; Kehinde, O. An established preadipose cell line and its differentiation in culture II. Factors affecting the adipose conversion. Cell 5:19–27; 1975.
Grimaud, J. A.; Borojevic, R. Myofibroblasts in hepatic schistosomal fibrosis. Experientia 33:890–892; 1977.
Guaragna, R. M.; Moraes, V. L.; Borojevic, R. Liver connective tissue cells: in vitro induction of the fat-storing phenotype. Arq. Biol. Tecnol. 32:36; 1989.
Hausman, G. J.; Novakofsky, J.; Martin, R. J., et al. The histochemistry of developing adipocytes in primary stromalvascular cultures of rat adipose tissue. Histochemistry 80:353–358; 1984.
Hirsch, J. G.; Fedorko, M. E. Ultrastructure of human leukocytes after simultaneous fixation with glutaraldehyde and osmium tetroxide and “post-fixation” in uranyl acetate. J. Cell Biol. 38:615–623; 1968.
Iyama, K.; Ohzono, K.; Usuku, G. Electron microscope studies on the genesis of white adipocytes: differentiation of immature pericytes into adipocytes in transplanted tissue. Virchows Arch. B Cell Pathol. 31:143–155; 1979.
Kawase, T.; Shiratori, Y.; Sugimoto, T. Collagen production by rat liver fat-storing cells in primary culture. Exp. Cell Biol. 54:183–192; 1986.
Knook, D. L.; Seffelaar, A. M. Fat-storing cells of the liver. Their isolation and purification. Exp. Cell Res. 139:486–471; 1982.
Leo, M. A.; Mak, K. M.; Savolainen, E. R., et al. Isolation and culture of myofibroblasts from rat liver Proc. Soc. Exp. Biol. Med. 180:382–391; 1985.
Mak, K.; Leo, M. A.; Lieber, C. S. Alcoholic liver injury in baboons: transformation of lipocytes to transitional cells. Gastroenterology 87:188–200; 1984.
Margis, R.; Borojevic, R. Retinoid-mediated induction of the fatstoring phenotype in a liver connective tissue cell line (GRX). Biochim. Biophys. Acta 1011:1–5; 1989.
Margis, M. P.; Domont, G.; Borojevic, R. Collagen biosynthesis during the adipose conversion in the GRX cell line. Arq. Biol. Tecnol. 31:27; 1988.
McPhail, L. C.; Clayton, C. C.; Snyderman, R. A potential second messenger role for unsaturated fatty acids: activation of Ca2+-dependent protein kinase. Science 224:622–625; 1984.
Minato, Y.; Hasumura, Y.; Takeuchi, J. The role of fat-storing cells in the Disse space fibrogenesis in alcoholic liver disease. Hepatology 4:559–566; 1983.
Monteiro, A. N. A.; Borojevic R. In vitro formation of fibrous septa by liver connective tissue cells. In Vitro Cell. Dev. Biol. 23:10–14; 1987.
Mourao, P. A. S.; Luz, M.; Borojevic, R. Sulfated glycosaminoglycans synthesis by human smooth muscle cells isolated from different organs. Biochim. Biophys Acta 881:321–329; 1986.
Nakano, M.; Worner, T. M.; Lieber, C. S. Perivenular fibrosis in alcoholic liver injury: ultrastructural and histologic progression. Gastroenterology 83:777–785; 1982.
Novikoff, A. B.; Novikoff, H. M.; Rosen, O. M., et al. Organelle relationship in cultured 3T3-Ll preadipocytes. J. Cell Biol. 87:180–196; 1980.
Rieder, H.; Teutsch, H. F.; Sasse, D. NADP-dependent dehydrogenases in rat liver parenchyma. Histochemistry 56:283–298; 1978.
Rudolph, R.; McLure, W. J.; Woodward, M. Contractible fibroblasts in chronic alcoholic cirrhosis. Gastroenterology 76:704–709; 1979.
Savolainen, E. R.; Leo, M. A.; Timpl, R., et al. Acetaldehyde and lactate stimulate collagen synthesis of cultured baboon liver myofibroblasts. Gastroenterology 87:777–787; 1984.
Sedmak, J. J.; Grossberg, S. E. A rapid sensitive and versatile assay for protein using Coomassie brilliant blue G250. Anal. Biochem. 79:544–552; 1977.
Spiegelman, B. V. M.; Green, H. Control of specific protein biosynthesis during the adipose conversion of 3T3 cells. J. Biol. Chem. 255:8811–8818; 1980.
Tanuma, Y.; Ito, T.; Shibasaki, S. Further electron microscope studies on the human hepatic sinusoidal wall with special reference to the fat-storing cells. Arch. Histol. Jpn. 45:263–274; 1982.
Verrando, P.; Negrel, R.; Grimaldi, P., et al. Differentiation of ob17 preadipocytes to adipocytes. Triggering effects of clofenopate and indomethacin. Biochim. Biophys. Acta 663:255–265; 1981.
Voss, B.; Rauterberg, J.; Pott, G., et al. Nonparenchymal cells cultivated from explants of fibrotic livers resemble endothelial and smooth muscle cells from vessel walls. Hepatology 2:19–28; 1982.
Williams, I. H.; Polakis, S.E. Differentiation of 3T3-Ll fibroblasts to adipocytes. The effect of indomethacin, prostaglandin E1 and cyclic AMP on the process of differentiation. Biochem. Biophys. Res. Comm. 77:175–186; 1977.
Yamamoto, K.; Ogawa, K. Fine structure and cytochemistry of lysosomes in the Ito cells of the liver. Cell Tissue Res. 233:45–57; 1983.
Zerbe, O.; Gressner, A. M. Proliferation of fat-storing cells is stimulated by secretions of Kupffer cells from normal and injured liver. Exp. Mol. Pathol. 49:87–101; 1988.
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This work was supported by FINEP and CNPq (Brazil).
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Borojevic, R., Guaragna, R.M., Margis, R. et al. In vitro induction of the fat-storing phenotype in a liver connective tissue cell line-GRX. In Vitro Cell Dev Biol 26, 361–368 (1990). https://doi.org/10.1007/BF02623827
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DOI: https://doi.org/10.1007/BF02623827