Skip to main content
SpringerLink
Log in

Ultrastructure of cultured hepatocytes from fat- and cholesterol-fed rats

  • Published:
Virchows Archiv B Cell Pathology Including Molecular Pathology

Summary

Hepatocytes were enzymatically dissociated from the livers of rats maintained for eight weeks on diets of (1) standard rat chow, (2) chow plus 20% olive oil, and (3) chow plus 20% olive oil and 2% cholesterol. Cells were cultured for up to one week and characterized by phase contrast light microscopy and transmission electron microscopy. Cultured hepatocytes derived from the cholesterol-fed rats (3) were distinguishable by phase contrast light microscopy from those derived from chow-or chow plus olive oil-fed (1 and 2) cells, since they contained numerous refractile cytoplasmic structures. Transmission electron microscopy demonstrated that these structures were lipid droplets. Hepatocytes were observed to accumulate non-membrane bound cytoplasmic lipid droplets in the order control-fed <olive oil-fed <cholesterol-fed. In addition to varying quantitatively among the feeding groups, lipid droplets also varied in size and osmiophilia as a function of the feeding regimen. Other than the differential accumulation of lipid droplets, the ultrastructure of the cultured hepatocytes was similar among cells derived from the various feeding groups.

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

Access this article

Log in via an institution

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

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Baglio CG, Farber E (1965) Reversal by adenine of the ethionine induced lipid accumulation in the endoplasmic reticulum of the rat liver. J Cell Biol 27:591–601

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Bernaert D, Wanson JC, Drochmans P, Popowski A (1977) Effect of insulin on ultrastructure and glycogenesis in primary cultures of adult rat hepatocytes. J Cell Biol 74:878–900

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Berry MN, Friend DS (1970) High-yield preparation of isolated rat liver parenchymal cells. J Cell Biol 43:506–520

    Article  Google Scholar 

  • 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

    Article  CAS  PubMed  Google Scholar 

  • Capuzzi DM, Margolis S (1971) Metabolic studies in isolated rat liver cells. II. Biosynthesis of serum low density lipoproteins and its regulation. Lipids 6:609–616

    CAS  PubMed  Google Scholar 

  • Chapman GS, Jones AL, Meyer UA, Bissell DM (1973) Parenchymal cells from adult rat liver in nonproliferating monolayer culture. II. Ultrastructural studies. J Cell Biol 59:735–747

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Claude A (1970) Growth and differentiation of cytoplasmic membranes in the course of lipoprotein granule synthesis. J Cell Biol 47:745–766

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Coleman R, Bell RM (1978) Evidence that biosynthesis of phosphatidylethanolamine, phosphatidylcholine, and triacylglycerol occurs on the cytoplasmic side of microsomal vesicles. J Cell Biol 76:245–253

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Davis RA, Showalter R, Kern F (1978) Reversal by triton WR-1339 of ethynyloestradiol-induced hepatic cholesterol esterification. Biochem J 174:45–51

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Davis RA, Engelhorn SC, Pangburn SH, Weinstein DB (1979a) Very low density lipoprotein synthesis and secretion by cultured rat hepatocytes. J Biol Chem 254:2010–2016

    CAS  PubMed  Google Scholar 

  • Davis RA, Englehorn SC, Weinstein DB, Steinberg D (1979b) Very low density lipoprotein secretion by cultured rat hepatocytes: Inhibition by albumin and other macromolecules. J Biol Chem 255:2039–2045

    Google Scholar 

  • Davis RA, McNeal M, Moses RL (1982) Intrahepatic assembly of very low density lipoprotein: Competition by cholesterol esters for the hydrophobic core. J Biol Chem 257:2634–2640

    CAS  PubMed  Google Scholar 

  • DeBrabander M, Wanson JC, Mosselmans R, Geuens G, Drochmans P (1978) Effects of anti-microtubular compounds on monolayer cultures of adult rat hepatocytes. Biol Cellul 31:127–140

    Google Scholar 

  • Deckelbaum RJ, Shipley GG, Small DM, Lee RS, George PK (1975) Thermal transitions in human plasma low density lipoproteins. Science 190:392–394

    Article  CAS  PubMed  Google Scholar 

  • Drevon CA, Weinstein DB, Steinberg D (1980) Regulation of cholesterol esterification and biosynthesis in monolayer cultures of normal rat hepatocytes. J Biol Chem 255:9128–9137

    CAS  PubMed  Google Scholar 

  • Drochmans P, Wanson JC, Mosselmans R (1975) Isolation and subfractionatin on ficol gradients of adult rat hepatocytes. J Cell Biol 66:1–22

    Article  CAS  PubMed  Google Scholar 

  • Elshourbagy NA, Barwick JL, Guzelian PS (1981) Induction of cytochrome P-450 by pregnenolone-16-carbonitrile in primary monolayer cultures of adult rat hepatocytes and in cell-free translation system. J Biol Chem 256:6060–6068

    CAS  PubMed  Google Scholar 

  • French SW, Davies PL (1975) Ultrastructural localization of actin like filaments in rat hepatocytes. Gastroent 68:765–774

    CAS  Google Scholar 

  • Goldstein AL, Johnson PR (1980) Primary cultures of fetal hepatocytes from the genetically obese Zucker rat: Characterization and total lipogenesis, in Vitro 16:288–296

    Article  CAS  Google Scholar 

  • Haagsman HP, Van Golde MG (1981) Synthesis and secretion of very low density lipoproteins by isolated rat hepatocytes in suspension: Role of diacylglycerol acyltransferase. Arch Biochem Biophys 208:395–402

    Article  CAS  PubMed  Google Scholar 

  • Hamilton RL, Regen DN, Gray ME, LeQuire VS (1967) Lipid transport in liver. I. Electron microscopic identification of very low density lipoproteins in perfused rat liver. Lab Invest 16:305–319

    CAS  PubMed  Google Scholar 

  • Iseri OA, Lieber CS, Gottleib LS (1966) The ultrastructure of fatty liver induced by prolonged ethanol ingestion. Am J Pathol 48:535–55

    PubMed Central  CAS  PubMed  Google Scholar 

  • Iype PT (1971) Cultures from adult rat liver cells. I. Establishment of monolayer cultures from normal liver. J Cell Phys 78:281–288

    Article  CAS  Google Scholar 

  • Jones AL, Fawcett DW (1966) Hypertrophy of the agranular endoplasmic reticulum in hamster liver induced by phenobarbital. J Histochem Cytochem 14:215–232

    Article  CAS  PubMed  Google Scholar 

  • Jones AL, Ruderman NB, Herrera MG (1967) Electron microscopic and biochemical study of lipoprotein synthesis in isolated perfused rat liver. J Lip Res 8:429–446

    CAS  Google Scholar 

  • Kroon PA, (1981) The order-disorder transition of the core cholesteryl esters of human plasma low denisty lipoprotein. J Biol Chem 256:5332–5238

    CAS  PubMed  Google Scholar 

  • Lee SS, Ho KJ (1975) Cholesterol fatty liver. Arch Pathol 99:301–306

    CAS  PubMed  Google Scholar 

  • Leffert HL, Paul D (1973) Serum dependent growth of primary cultured differentiated fetal hepatocytes in arginine deficient medium. J Cell Phys 81:113–124

    Article  CAS  Google Scholar 

  • Lieber CS, DeCarli LM (1970) Quantitative relationships between the amount of dietary fat and severity of alcoholic fatty liver. Am J Clin Nutr 23:474–478

    CAS  PubMed  Google Scholar 

  • Loud AV (1968) A quantitative sterological description of the ultrastructure of normal rat liver parenchymal cells. J Cell Biol 37:27–46

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Mahley RW, Gray ME, Hamilton RL, Lequire VS (1968) Lipid transport in liver. II. Electron microscopic and biochemical studies of alterations in lipoprotein transport induced by cortisone in the rabbit. Lab Invest 19:358–369

    CAS  PubMed  Google Scholar 

  • Mahley RW, Hamilton RL, Lequire VS (1969) Characterization of lipoprotein particles isolated from the Golgi apparatus of rat liver. J Lip Res 10:433–439

    CAS  Google Scholar 

  • Montesano R, Mira-Moser F, Stefan Y, Perrelet A, Orci L (1978) Tight junctions in fetal rat liver expiants grown in vitro. J Ultrastruct Res 64:182–190

    Article  CAS  PubMed  Google Scholar 

  • Morre DJ, Merritt WD, Lembi CA (1971) Connections between mitochondria and endoplasmic reticulum in rat liver and onion stem. Protoplasma 73:43–49

    Article  CAS  PubMed  Google Scholar 

  • Moses RL, Davis RA (1980) Cell morphology and very low density lipoprotein secretion by cultured hepatocytes derived from fat-fed rats. J Cell Biol 87:305a

    Google Scholar 

  • Moses RL, Davis RA (1981) Morphology of cultured hepatocytes from fat-fed rats. Anat Rec 199:177A

    Article  Google Scholar 

  • Novikoff AB, Roheim PS, Quintana N (1966) Changes in rat liver cells induced by orotic acid feeding. Lab Invest 15:27–49

    CAS  Google Scholar 

  • Ontko JA (1973) Effects of ethanol on the metabolism of free fatty acids in isolated liver cells. J Lip Res 14:78–86

    CAS  Google Scholar 

  • Parks HF (1967) An experimental study of microscopic and submicroscopic lipid inclusions in hepatic cells of the mouse. Am J Anat 120:253–280

    Article  Google Scholar 

  • Ridout JH, Lucas CC, Patterson JN, Best CH (1954) Changes in chemical composition during the development of “cholesterol fatty livers”. Biochem J 58:297–301

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Schanne FAX, Zuker AH, Farber JL (1981) Alcohol-dependent liver cell necrosis in vitro: A new model. Science (US) 212:338–340

    CAS  Google Scholar 

  • Stein O, Stein Y (1965) Fine structure of the ethanol induced fatty liver in the rat. Isr J Med Sci 1:378–388

    Google Scholar 

  • Stein O, Stein Y (1966) Electron microscopic autoradiography of3H glycerol labelled lipid in ethanol induced fatty liver. Exp Cell Res 42:198–201

    Article  CAS  PubMed  Google Scholar 

  • Stein O, Stein Y (1967) Lipid synthesis, intracellular transport, storage, and secretion. I. Electron microscopic radioautographic study of liver after injection of tritrated palmitate or glycerol in fasted and ethanol treated rats. J Cell Biol 33:319–339

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  • Tarlow DM, Watkins PA, Reed RE, Miller RS, Zwergel EE, Lane MD (1977) Lipogenesis and the synthesis and secretion of very low density lipoprotein by avian liver cells in nonproliferating monolayer culture. J Cell Biol 73:332–353

    Article  CAS  PubMed  Google Scholar 

  • Wanson JC, Drochmans P, Mosselmans R, Ronveaux MF (1977) Adult heaptocytes in primary monolayer culture. J Cell Biol 74:858–877

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Anatomy, Department of Physiology Louisiana State University Medical Center, 70119, New Orleans, LA, USA

    R. L. Moses & Joseph B. Delcarpio

  2. Cell Biology Unit, Department of Physiology Louisiana State University Medical Center, 70119, New Orleans, LA, USA

    Monica Malone McNeal & Roger A. Davis

Authors
  1. R. L. Moses
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Joseph B. Delcarpio
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Monica Malone McNeal
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Roger A. Davis
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

Supported by a grant from the E.G. Schlieder Educational Foundation (R.L.M.) and NIH grants HL 23895 and HL25596, (R.A.D.). R.A.D. is an Established Investigator of the American Heart Association. The authors thank Ms. Toni Santee for typing and Ms. Carla Moses for editing this manuscript

Rights and permissions

Reprints and permissions

About this article

Cite this article

Moses, R.L., Delcarpio, J.B., McNeal, M.M. et al. Ultrastructure of cultured hepatocytes from fat- and cholesterol-fed rats. Virchows Archiv B Cell Pathol 41, 95–106 (1982). https://doi.org/10.1007/BF02890274

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF02890274

Key words

Search

Navigation