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Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1985 Dec;82(24):8681–8685. doi: 10.1073/pnas.82.24.8681

Hepatic lipocytes: the principal collagen-producing cells of normal rat liver.

S L Friedman, F J Roll, J Boyles, D M Bissell
PMCID: PMC391500  PMID: 3909149

Abstract

Hepatic lipocytes were isolated from normal rat liver and established in culture. A virtually pure isolate was obtained by fractionating enzymatically digested liver on a discontinuous gradient of arabinogalactan. Isolated cells displayed prominent rough endoplasmic reticulum and typical cytoplasmic droplets containing vitamin A. Lipocytes in primary culture were shown by immunofluorescence to secrete collagen types I, III, and IV and also laminin. Immunoassay of culture media showed that lipocytes during the first week in culture secrete type I (72.1-86.2% of total measured soluble collagen), type III (2.6-7.2%), and type IV (11.2-29.7%) collagens. Five percent of total secreted protein was collagen compared with 0.2% in similarly cultured hepatocytes and 1.7% in sinusoidal endothelial cells, as measured by the production of peptide-bound [3H]hydroxyproline in cells incubated with [3H]proline. The calculated amount of collagen synthesized by lipocytes per microgram of cellular DNA was 10-fold greater than that produced by hepatocytes and over 20-fold greater than that produced by endothelial cells. The findings indicate that collagen synthesis and secretion are specialized functions of hepatic lipocytes, and that, in cells from normal liver, this represents production primarily of type I collagen. The phenotypic resemblance of these cells to fibroblasts supports the hypothesis that lipocytes are a major source of collagen in pathologic fibrosis and may be precursors of the fibroblast-like cells observed in liver injury.

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Selected References

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  1. Airhart J., Kelley J., Brayden J. E., Low R. B., Stirewalt W. S. An ultramicro method of amino acid analysis: application to studies of protein metabolism in cultured cells. Anal Biochem. 1979 Jul 1;96(1):45–55. doi: 10.1016/0003-2697(79)90552-9. [DOI] [PubMed] [Google Scholar]
  2. Bienkowski R. S., Cowan M. J., McDonald J. A., Crystal R. G. Degradation of newly synthesized collagen. J Biol Chem. 1978 Jun 25;253(12):4356–4363. [PubMed] [Google Scholar]
  3. Bissell D. M., Guzelian P. S. Degradation of endogenous hepatic heme by pathways not yielding carbon monoxide. Studies in normal rat liver and in primary hepatocyte culture. J Clin Invest. 1980 May;65(5):1135–1140. doi: 10.1172/JCI109767. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Blomhoff R., Holte K., Naess L., Berg T. Newly administered [3H]retinol is transferred from hepatocytes to stellate cells in liver for storage. Exp Cell Res. 1984 Jan;150(1):186–193. doi: 10.1016/0014-4827(84)90713-4. [DOI] [PubMed] [Google Scholar]
  5. Clément B., Emonard H., Rissel M., Druguet M., Grimaud J. A., Herbage D., Bourel M., Guillouzo A. Cellular origin of collagen and fibronectin in the liver. Cell Mol Biol. 1984;30(5):489–496. [PubMed] [Google Scholar]
  6. Corash L., Tan H., Gralnick H. R. Heterogeneity of human whole blood platelet subpopulations. I. Relationship between buoyant density, cell volume, and ultrastructure. Blood. 1977 Jan;49(1):71–87. [PubMed] [Google Scholar]
  7. Diegelmann R. F., Guzelian P. S., Gay R., Gay S. Collagen formation by the hepatocyte in primary monolayer culture and in vivo. Science. 1983 Mar 18;219(4590):1343–1345. doi: 10.1126/science.6828863. [DOI] [PubMed] [Google Scholar]
  8. Diegelmann R. F., Peterkofsky B. Inhibition of collagen secretion from bone and cultured fibroblasts by microtubular disruptive drugs. Proc Natl Acad Sci U S A. 1972 Apr;69(4):892–896. doi: 10.1073/pnas.69.4.892. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Fahimi H. D. The fine structural localization of endogenous and exogenous peroxidase activity in Kupffer cells of rat liver. J Cell Biol. 1970 Oct;47(1):247–262. doi: 10.1083/jcb.47.1.247. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Fleischmajer R., Perlish J. S., Krieg T., Timpl R. Variability in collagen and fibronectin synthesis by scleroderma fibroblasts in primary culture. J Invest Dermatol. 1981 May;76(5):400–403. doi: 10.1111/1523-1747.ep12520933. [DOI] [PubMed] [Google Scholar]
  11. GIRARDI A. J., MCMICHAEL H., Jr, HENLE W. The use of HeLa cells in suspension for the quantitative study of virus propagation. Virology. 1956 Aug;2(4):532–544. doi: 10.1016/0042-6822(56)90009-5. [DOI] [PubMed] [Google Scholar]
  12. Green H., Todaro G. J., Goldberg B. Collagen synthesis in fibroblasts transformed by oncogenic viruses. Nature. 1966 Feb 26;209(5026):916–917. doi: 10.1038/209916a0. [DOI] [PubMed] [Google Scholar]
  13. Herrmann H., Dessau W., Fessler L. I., von der Mark K. Synthesis of types I, III and AB2 collagen by chick tendon fibroblasts in vitro. Eur J Biochem. 1980 Mar;105(1):63–74. doi: 10.1111/j.1432-1033.1980.tb04474.x. [DOI] [PubMed] [Google Scholar]
  14. Irving M. G., Roll F. J., Huang S., Bissell D. M. Characterization and culture of sinusoidal endothelium from normal rat liver: lipoprotein uptake and collagen phenotype. Gastroenterology. 1984 Dec;87(6):1233–1247. [PubMed] [Google Scholar]
  15. Kent G., Gay S., Inouye T., Bahu R., Minick O. T., Popper H. Vitamin A-containing lipocytes and formation of type III collagen in liver injury. Proc Natl Acad Sci U S A. 1976 Oct;73(10):3719–3722. doi: 10.1073/pnas.73.10.3719. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Knook D. L., Seffelaar A. M., de Leeuw A. M. Fat-storing cells of the rat liver. Their isolation and purification. Exp Cell Res. 1982 Jun;139(2):468–471. doi: 10.1016/0014-4827(82)90283-x. [DOI] [PubMed] [Google Scholar]
  17. Labarca C., Paigen K. A simple, rapid, and sensitive DNA assay procedure. Anal Biochem. 1980 Mar 1;102(2):344–352. doi: 10.1016/0003-2697(80)90165-7. [DOI] [PubMed] [Google Scholar]
  18. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  19. Lazarides E. Intermediate filaments as mechanical integrators of cellular space. Nature. 1980 Jan 17;283(5744):249–256. doi: 10.1038/283249a0. [DOI] [PubMed] [Google Scholar]
  20. Madri J. A., Barwick K. W. Use of avidin-biotin complex in an ELISA system: a quantitative comparison with two other immunoperoxidase detection systems using keratin antisera. Lab Invest. 1983 Jan;48(1):98–107. [PubMed] [Google Scholar]
  21. Mayne R., Vail M. S., Miller E. J. Characterization of the collagen chains synthesized by cultured smooth muscle cells derived from rhesus monkey thoracic aorta. Biochemistry. 1978 Feb 7;17(3):446–452. doi: 10.1021/bi00596a011. [DOI] [PubMed] [Google Scholar]
  22. Minato Y., Hasumura Y., Takeuchi J. The role of fat-storing cells in Disse space fibrogenesis in alcoholic liver disease. Hepatology. 1983 Jul-Aug;3(4):559–566. doi: 10.1002/hep.1840030414. [DOI] [PubMed] [Google Scholar]
  23. Okanoue T., Burbige E. J., French S. W. The role of the Ito cell in perivenular and intralobular fibrosis in alcoholic hepatitis. Arch Pathol Lab Med. 1983 Sep;107(9):459–463. [PubMed] [Google Scholar]
  24. Rennard S. I., Berg R., Martin G. R., Foidart J. M., Robey P. G. Enzyme-linked immunoassay (ELISA) for connective tissue components. Anal Biochem. 1980 May 1;104(1):205–214. doi: 10.1016/0003-2697(80)90300-0. [DOI] [PubMed] [Google Scholar]
  25. Roll F. J., Madri J. A., Albert J., Furthmayr H. Codistribution of collagen types IV and AB2 in basement membranes and mesangium of the kidney. an immunoferritin study of ultrathin frozen sections. J Cell Biol. 1980 Jun;85(3):597–616. doi: 10.1083/jcb.85.3.597. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Roll F. J., Madri J. A., Furthmayr H. A new method of iodinating collagens for use in radioimmunoassay. Anal Biochem. 1979 Jul 15;96(2):489–499. doi: 10.1016/0003-2697(79)90611-0. [DOI] [PubMed] [Google Scholar]
  27. Smith B. D., McKenney K. H., Lustberg T. J. Characterization of collagen precursors found in rat skin and rat bone. Biochemistry. 1977 Jun 28;16(13):2980–2985. doi: 10.1021/bi00632a027. [DOI] [PubMed] [Google Scholar]
  28. Smith F. R., Goodman D. S. Vitamin A transport in human vitamin A toxicity. N Engl J Med. 1976 Apr 8;294(15):805–808. doi: 10.1056/NEJM197604082941503. [DOI] [PubMed] [Google Scholar]
  29. Tseng S. C., Lee P. C., Ells P. F., Bissell D. M., Smuckler E. A., Stern R. Collagen production by rat hepatocytes and sinusoidal cells in primary monolayer culture. Hepatology. 1982 Jan-Feb;2(1):13–18. doi: 10.1002/hep.1840020104. [DOI] [PubMed] [Google Scholar]
  30. Wake K. Perisinusoidal stellate cells (fat-storing cells, interstitial cells, lipocytes), their related structure in and around the liver sinusoids, and vitamin A-storing cells in extrahepatic organs. Int Rev Cytol. 1980;66:303–353. doi: 10.1016/s0074-7696(08)61977-4. [DOI] [PubMed] [Google Scholar]
  31. Weibel E. R., Stäubli W., Gnägi H. R., Hess F. A. Correlated morphometric and biochemical studies on the liver cell. I. Morphometric model, stereologic methods, and normal morphometric data for rat liver. J Cell Biol. 1969 Jul;42(1):68–91. doi: 10.1083/jcb.42.1.68. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Wells A. F., Miller C. E., Nadel M. K. Rapid fluorescein and protein assay method for fluorescent-antibody conjugates. Appl Microbiol. 1966 Mar;14(2):271–275. doi: 10.1128/am.14.2.271-275.1966. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Yokoi Y., Namihisa T., Kuroda H., Komatsu I., Miyazaki A., Watanabe S., Usui K. Immunocytochemical detection of desmin in fat-storing cells (Ito cells). Hepatology. 1984 Jul-Aug;4(4):709–714. doi: 10.1002/hep.1840040425. [DOI] [PubMed] [Google Scholar]

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