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J Cell Biol. 1969 Dec 1; 43(3): 506–520.
PMCID: PMC2107801


A Biochemical and Fine Structural Study


A new technique employing continuous recirculating perfusion of the rat liver in situ, shaking of the liver in buffer in vitro, and filtration of the tissue through nylon mesh, results in the conversion of about 50% of the liver into intact, isolated parenchymal cells. The perfusion media consist of: (a) calcium-free Hanks' solution containing 0.05% collagenase and 0.10% hyaluronidase, and (b) magnesium and calcium-free Hanks' solution containing 2 mM ethylenediaminetetraacetate. Biochemical and morphologic studies indicate that the isolated cells are viable. They respire in a medium containing calcium ions, synthesize glucose from lactate, are impermeable to inulin, do not stain with trypan blue, and retain their structural integrity. Electron microscopy of biopsies taken during and after perfusion reveals that desmosomes are quickly cleaved. Hemidesmosome-containing areas of the cell membrane invaginate and appear to pinch off and migrate centrally. Tight and gap junctions, however, persist on the intact, isolated cells, retaining small segments of cytoplasm from formerly apposing parenchymal cells. Cells which do not retain tight and gap junctions display swelling of Golgi vacuoles and vacuoles in the peripheral cytoplasm. Cytoplasmic vacuolization in a small percentage of cells and potassium loss are the only indications of cell injury detected. By other parameters measured, the isolated cells are comparable to normal hepatic parenchymal cells in situ in appearance and function.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • ANDERSON NG. The mass isolation of whole cells from rat liver. Science. 1953 Jun 5;117(3049):627–628. [PubMed]
  • KALTENBACH JP. The preparation and utilization of whole cell suspensions obtained from solid mammalian tissues. Exp Cell Res. 1954 Nov;7(2):568–571. [PubMed]
  • LONGMUIR IS, REES WAP. Preparation of cell suspensions from rat livers. Nature. 1956 May 26;177(4517):997–997. [PubMed]
  • BRANSTER MV, MORTON RK. Isolation of intact liver cells. Nature. 1957 Dec 7;180(4597):1283–1284. [PubMed]
  • JACOB ST, BHARGAVA PM. A new method for the preparation of liver cell suspensions. Exp Cell Res. 1962 Sep;27:453–467. [PubMed]
  • LAWS JO, STRICKLAND LH. Metabolism of isolated liver cells. Nature. 1956 Aug 11;178(4528):309–310. [PubMed]
  • Howard RB, Christensen AK, Gibbs FA, Pesch LA. The enzymatic preparation of isolated intact parenchymal cells from rat liver. J Cell Biol. 1967 Dec;35(3):675–684. [PMC free article] [PubMed]
  • Howard RB, Pesch LA. Respiratory activity of intact, isolated parenchymal cells from rat liver. J Biol Chem. 1968 Jun 10;243(11):3105–3109. [PubMed]
  • MORTIMORE GE. Effect of insulin on potassium transfer in isolated rat liver. Am J Physiol. 1961 Jun;200:1315–1319. [PubMed]
  • HANKS JH, WALLACE RE. Relation of oxygen and temperature in the preservation of tissues by refrigeration. Proc Soc Exp Biol Med. 1949 Jun;71(2):196–200. [PubMed]
  • LUFT JH. Improvements in epoxy resin embedding methods. J Biophys Biochem Cytol. 1961 Feb;9:409–414. [PMC free article] [PubMed]
  • NOVIKOFF AB, GOLDFISCHER S. Nucleosidediphosphatase activity in the Golgi apparatus and its usefulness for cytological studies. Proc Natl Acad Sci U S A. 1961 Jun 15;47:802–810. [PMC free article] [PubMed]
  • SCHMIDT FH. [Enzymatic determination of glucose and fructose simultaneously]. Klin Wochenschr. 1961 Dec 1;39:1244–1247. [PubMed]
  • WROBLEWSKI F, LADUE JS. Lactic dehydrogenase activity in blood. Proc Soc Exp Biol Med. 1955 Oct;90(1):210–213. [PubMed]
  • HENLEY KS, POLLARD HM. A new method for the determination of glutamic oxalacetic and glutamic pyruvic transaminase in plasma. J Lab Clin Med. 1955 Nov;46(5):785–789. [PubMed]
  • KULKA RG. Colorimetric estimation of ketopentoses and ketohexoses. Biochem J. 1956 Aug;63(4):542–548. [PMC free article] [PubMed]
  • CHANEY AL, MARBACH EP. Modified reagents for determination of urea and ammonia. Clin Chem. 1962 Apr;8:130–132. [PubMed]
  • LOWRY OH, ROSEBROUGH NJ, FARR AL, RANDALL RJ. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed]
  • WERKHEISER WC, BARTLEY W. The study of steady-state concentrations of internal solutes of mitochondria by rapid centrifugal transfer to a fixation medium. Biochem J. 1957 May;66(1):79–91. [PMC free article] [PubMed]
  • Heath T, Wissig SL. Fine structure of the surface of mouse hepatic cells. Am J Anat. 1966 Jul;119(1):97–127. [PubMed]
  • Revel JP, Karnovsky MJ. Hexagonal array of subunits in intercellular junctions of the mouse heart and liver. J Cell Biol. 1967 Jun;33(3):C7–C12. [PMC free article] [PubMed]
  • FARQUHAR MG, PALADE GE. Junctional complexes in various epithelia. J Cell Biol. 1963 May;17:375–412. [PMC free article] [PubMed]
  • FAWCETT DW. Observations on the cytology and electron microscopy of hepatic cells. J Natl Cancer Inst. 1955 Apr;15(5 Suppl):1475–1503. [PubMed]
  • Jones AL, Ruderman NB, Herrera MG. An electron microscopic study of lipoprotein production and release by the isolated perfused rat liver. Proc Soc Exp Biol Med. 1966 Oct;123(1):4–9. [PubMed]
  • HENLEY KS, WIGGINS HS, POLLARD HM, DULLAERT E. The transaminase content of parenchymatous liver cells. Gastroenterology. 1959 Jan;36(1):1–6. [PubMed]
  • ZIMMERMAN M, DEVLIN TM, PRUSS MP. Anaerobic glycolysis of dispersed cell suspensions from normal and malignant tissues. Nature. 1960 Jan 30;185:315–316. [PubMed]
  • CONWAY EJ, DOWNEY M. An outer metabolic region of the yeast cell. Biochem J. 1950 Sep;47(3):347–355. [PMC free article] [PubMed]
  • Hems R, Ross BD, Berry MN, Krebs HA. Gluconeogenesis in the perfused rat liver. Biochem J. 1966 Nov;101(2):284–292. [PMC free article] [PubMed]
  • MILLER LL, BLY CG, WATSON ML, BALE WF. The dominant role of the liver in plasma protein synthesis; a direct study of the isolated perfused rat liver with the aid of lysine-epsilon-C14. J Exp Med. 1951 Nov;94(5):431–453. [PMC free article] [PubMed]
  • Jones AL, Ruderman NB, Herrera MG. Electron microscopic and biochemical study of lipoprotein synthesis in the isolated perfused rat liver. J Lipid Res. 1967 Sep;8(5):429–446. [PubMed]
  • Jacob ST, Bhargava PM. Effect of chloramphenicol on ribonucleic acid synthesis in liver cells in suspension. Biochem J. 1965 Oct;97(1):67–73. [PMC free article] [PubMed]
  • Ontko JA. Chylomicron, free fatty acid and ketone body metabolism of isolated liver cells and liver homogenates. Biochim Biophys Acta. 1967 Feb 14;137(1):13–22. [PubMed]
  • Hamilton RL, Regen DM, Gray ME, LeQuire VS. Lipid transport in liver. I. Electron microscopic identification of very low density lipoproteins in perfused rat liver. Lab Invest. 1967 Feb;16(2):305–319. [PubMed]
  • Overton J. The fat of desmosomes in trypsinized tissue. J Exp Zool. 1968 Jun;168(2):203–214. [PubMed]

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