Isolation and characterization of Kupffer and endothelial cells from the rat liver

Knook, D.L.; Blansjaar, N.; Sleyster, E.Ch.

doi:10.1016/0014-4827(77)90011-8

Cited by 244 publications

(111 citation statements)

References 31 publications

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“…The viability äs checked by trypan blue exclusion was greater than 90%. Identification and assessment of the purity of the Kupffer cells were performed by light-and electron microscopy (38), peroxidase staining, by phagocytosis of latex beads (LB 11 polystyrene beads, mean diameter 1.1 ; Sigma Chem. Co., St. Louis, USA) for 60 min at 37 °C (23), and by demonstration of positive immunofluorescence staining for vimentin and negative staining for desmin (39).…”

Section: Methodsmentioning

confidence: 99%

Comparison of Sulphated Glycosaminoglycan and Hyaluronate Synthesis and Secretion in Cultured Hepatocytes, Fat Storing Cells, and Kupffer Cells

Gressner¹,

Schäfer²

1989

Clinical Chemistry and Laboratory Medicine

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Summary:The extracellular matrix of normal liver contains several types of proteoglycans including heparan sulphate, chondroitin sulphate isomers, dermatan sulphate, and the glycosaminoglycan, hyaluronic acid. In the present study both the synthesis and secretion äs well äs the pattern of radioactively labeled proteoglycans and hyaluronic acid of hepatocytes, fat-storing cells (Ito cells), and Kupffer cells maintained in monolayer cultures under mostly identical conditions were compared to assess their relative contribution to hepatic proteoglycan synthesis. Fat-storing cells were identified äs the main type of cell producing and secreting proteoglycan and hyaluronic acid. More than 70% of labeled proteoglycan and hyaluronic acid were secreted into the medium. Heparan sulphate is the main type of proteoglycan in hepatocytes, whereas in the medium of fat-storing cells, chondroitin sulphate and dermatan sulphate comprise the mäjor fractions. Hyaluronic acid was not detectable in hepatocyte cultures and found only in low amounts in the medium of Kupffer cells. The results point to a stringent quantitative and qualitative cellular compartmentation of proteoglycan synthesis in liver with fat-storing cells äs the most important cell type for matrix proteoglycan and hyaluronic acid production.

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Section: Methodsmentioning

confidence: 99%

Comparison of Sulphated Glycosaminoglycan and Hyaluronate Synthesis and Secretion in Cultured Hepatocytes, Fat Storing Cells, and Kupffer Cells

Gressner¹,

Schäfer²

1989

Clinical Chemistry and Laboratory Medicine

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“…[18][19][20][21][22][23] Mean relative purity of freshly isolated cells was 85% Ϯ 3%. Major contaminants were endothelial cells (10%) and Kupffer cells (3%).…”

mentioning

confidence: 99%

“…These concentrations of the cytokines have previously been shown to have no cytotoxic effects. 12,25 Time kinetic studies (6,10,14,16,18,20,24,30, and 36 hours) to investigate the influence of the incubation time on proliferation of HSC were also performed. Significant effects of TGF-␤ on apoptosis and proliferation could be observed after 14 hours of incubation and reached a maximum after 18 hours.…”

mentioning

confidence: 99%

Transforming growth factor β and tumor necrosis factor α inhibit both apoptosis and proliferation of activated rat hepatic stellate cells

et al. 1999

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Transforming growth factor ␤ (TGF-␤) as well as tumor necrosis factor ␣ (TNF-␣) gene expression are up-regulated in chronically inflamed liver. These cytokines were investigated for their influence on apoptosis and proliferation of activated hepatic stellate cells (HSCs). Spontaneous apoptosis in activated HSC was significantly down-regulated by 53% ؎ 8% (P F .01) under the influence of TGF-␤ and by 28% ؎ 2% (P F .05) under the influence of TNF-␣. TGF-␤ and TNF-␣ significantly reduced expression of CD95L in activated HSCs, whereas CD95 expression remained unchanged. Furthermore, HSC apoptosis induced by CD95-agonistic antibodies was reduced from 96% ؎ 2% to 51 ؎ 7% (P F .01) by TGF-␤, and from 96% ؎ 2% to 58 ؎ 2% (P F .01) by TNF-␣, suggesting that intracellular antiapoptotic mechanisms may also be activated by both cytokines. During activation, HSC cultures showed a reduced portion of cells in the G 0 /G 1 phase and a strong increment of G 2 -phase cells. This increment was significantly inhibited (G 1 arrest) by administration of TGF-␤ and/or TNF-␣ to activated cells. In liver sections of chronically damaged rat liver (CCl 4 model), using desmin and CD95L as markers for activated HSC, most of these cells did not show apoptotic signs (TUNEL-negative). Taken together, these findings indicate that TGF-␤ and/or TNF-␣ both inhibit proliferation and also apoptosis in activated HSC in vitro. Both processes seem to be linked to each other, and their inhibition could represent the mechanism responsible for prolonged survival of activated HSC in chronic liver damage in vivo. (HEPATOLOGY 1999;30:196-202.)

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“…Procedures developed for the preparation of highly purified Kupffer and endothelial cells [3,4] offer the possibility to determine the amount of ligand endocytosed by each of the cell types prior to their isolation. However, during the liver perfusion and subsesequent incubation of the cells at 37'C, degradation of the intracellular ligands can occur.…”

Section: Injection and Plasma Clearance Of The Ligandsmentioning

confidence: 99%

“…The main difference between this isolation method and the pronase E method in [3,4] is the reduced temperature during the entire isolation procedure. At 10°C pronase is apparently still able to digest the parenchymal cells, since the total sinusoidal cell suspension contained only l-2% parenchymal cells before elution.…”

Section: I Characterization Of the Cell Preparationsmentioning

confidence: 99%

Quantitative determination of in Vivo endocytosis by rat liver kupffer and endothelial cells facilitated by an improved cell isolation method

Dalen

Knook

1982

FEBS Letters

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Isolation and characterization of Kupffer and endothelial cells from the rat liver

Cited by 244 publications

References 31 publications

Comparison of Sulphated Glycosaminoglycan and Hyaluronate Synthesis and Secretion in Cultured Hepatocytes, Fat Storing Cells, and Kupffer Cells

Comparison of Sulphated Glycosaminoglycan and Hyaluronate Synthesis and Secretion in Cultured Hepatocytes, Fat Storing Cells, and Kupffer Cells

Transforming growth factor β and tumor necrosis factor α inhibit both apoptosis and proliferation of activated rat hepatic stellate cells

Quantitative determination of in Vivo endocytosis by rat liver kupffer and endothelial cells facilitated by an improved cell isolation method

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