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Liver sinusoidal endothelial cells represents an important blood clearance system in pigs.

Nedredal GI, Elvevold KH, Ytrebø LM, Olsen R, Revhaug A, Smedsrød B - Comp Hepatol (2003)

Bottom Line: Use of specific anti-Kupffer cell- and anti-desmin antibodies, combined with endocytosis of fluorescently labeled macromolecular soluble ligands indicated that the LSEC fraction contained 62 x 107 (sd 12 x 107) purified LSEC.Cultured LSEC avidly endocytosed ligands for the mannose and scavenger receptors.CONCLUSIONS: We show here for the first time that pig LSEC, similar to what has been found earlier in rat LSEC, represent an effective scavenger system for removal of macromolecular waste products from the circulation.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Digestive Surgery, University Hospital of Tromsø, 9038 Tromsø, Norway. Geir.Ivar.Nedredal@fagmed.uit.no

ABSTRACT
BACKGROUND: Numerous studies in rats and a few other mammalian species, including man, have shown that the sinusoidal cells constitute an important part of liver function. In the pig, however, which is frequently used in studies on liver transplantation and liver failure models, our knowledge about the function of hepatic sinusoidal cells is scarce. We have explored the scavenger function of pig liver sinusoidal endothelial cells (LSEC), a cell type that in other mammals performs vital elimination of an array of waste macromolecules from the circulation. RESULTS: 125I-macromolecules known to be cleared in the rat via the scavenger and mannose receptors were rapidly removed from the pig circulation, 50% of the injected dose being removed within the first 2-5 min following injection. Fluorescently labeled microbeads (2 &mgr;m in diameter) used to probe phagocytosis accumulated in Kupffer cells only, whereas fluorescently labeled soluble macromolecular ligands for the mannose and scavenger receptors were sequestered only by LSEC. Desmin-positive stellate cells accumulated no probes. Isolation of liver cells using collagenase perfusion through the portal vein, followed by various centrifugation protocols to separate the different liver cell populations yielded 280 x 107 (range 50-890 x 107) sinusoidal cells per liver (weight of liver 237.1 g (sd 43.6)). Use of specific anti-Kupffer cell- and anti-desmin antibodies, combined with endocytosis of fluorescently labeled macromolecular soluble ligands indicated that the LSEC fraction contained 62 x 107 (sd 12 x 107) purified LSEC. Cultured LSEC avidly endocytosed ligands for the mannose and scavenger receptors. CONCLUSIONS: We show here for the first time that pig LSEC, similar to what has been found earlier in rat LSEC, represent an effective scavenger system for removal of macromolecular waste products from the circulation.

No MeSH data available.


Related in: MedlinePlus

Fluorescent micrographs of cultured stellate cells and Kupffer cells. Cultures were prepared as described in Methods. The cultures were fixed in 4% paraformaldehyde, after 1 h of incubation. Micrographs of cultured stellate cells stained with monoclonal anti-desmin antibody (A) and cultured Kupffer cells stained with monoclonal anti-pig macrophage antibody (B). (Scale bars; 20 μm).
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Figure 7: Fluorescent micrographs of cultured stellate cells and Kupffer cells. Cultures were prepared as described in Methods. The cultures were fixed in 4% paraformaldehyde, after 1 h of incubation. Micrographs of cultured stellate cells stained with monoclonal anti-desmin antibody (A) and cultured Kupffer cells stained with monoclonal anti-pig macrophage antibody (B). (Scale bars; 20 μm).

Mentions: Cells, seeded on fibronectin-coated substrate, obtained from the elutriation fractions yielded LSEC cultures of varying purity (Table 3). We used in vivo (Fig. 6A) or in vitro administered FITC-FSA as a specific LSEC marker, positive reaction with anti-desmin antibodies as a specific marker of stellate cells (Fig. 7A), and a specific anti-pig macrophage antibody (Fig. 7B) or phagocytosis of TRITC-MDPP (Fig. 6B) as KC specific markers. Using these criteria, cultures resulting from elutriation fraction 1 were shown to contain 63.9% stellate cells; cultures established from fraction 2 contained 80.4% LSEC, and fractions 3 and 4 contained 66.2% and 61.0% LSEC. Cells that reacted with anti-pig-macrophage antibodies or phagocytosed TRITC-MDPP contained no FITC-FSA. Stellate cells were distinguished by immunolabeling with anti-desmin antibodies or by their content of characteristic autofluorescence from vitamin A droplets when irradiated with light of 328 nm of wavelength ([12]) (Fig. 6C).


Liver sinusoidal endothelial cells represents an important blood clearance system in pigs.

Nedredal GI, Elvevold KH, Ytrebø LM, Olsen R, Revhaug A, Smedsrød B - Comp Hepatol (2003)

Fluorescent micrographs of cultured stellate cells and Kupffer cells. Cultures were prepared as described in Methods. The cultures were fixed in 4% paraformaldehyde, after 1 h of incubation. Micrographs of cultured stellate cells stained with monoclonal anti-desmin antibody (A) and cultured Kupffer cells stained with monoclonal anti-pig macrophage antibody (B). (Scale bars; 20 μm).
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC149430&req=5

Figure 7: Fluorescent micrographs of cultured stellate cells and Kupffer cells. Cultures were prepared as described in Methods. The cultures were fixed in 4% paraformaldehyde, after 1 h of incubation. Micrographs of cultured stellate cells stained with monoclonal anti-desmin antibody (A) and cultured Kupffer cells stained with monoclonal anti-pig macrophage antibody (B). (Scale bars; 20 μm).
Mentions: Cells, seeded on fibronectin-coated substrate, obtained from the elutriation fractions yielded LSEC cultures of varying purity (Table 3). We used in vivo (Fig. 6A) or in vitro administered FITC-FSA as a specific LSEC marker, positive reaction with anti-desmin antibodies as a specific marker of stellate cells (Fig. 7A), and a specific anti-pig macrophage antibody (Fig. 7B) or phagocytosis of TRITC-MDPP (Fig. 6B) as KC specific markers. Using these criteria, cultures resulting from elutriation fraction 1 were shown to contain 63.9% stellate cells; cultures established from fraction 2 contained 80.4% LSEC, and fractions 3 and 4 contained 66.2% and 61.0% LSEC. Cells that reacted with anti-pig-macrophage antibodies or phagocytosed TRITC-MDPP contained no FITC-FSA. Stellate cells were distinguished by immunolabeling with anti-desmin antibodies or by their content of characteristic autofluorescence from vitamin A droplets when irradiated with light of 328 nm of wavelength ([12]) (Fig. 6C).

Bottom Line: Use of specific anti-Kupffer cell- and anti-desmin antibodies, combined with endocytosis of fluorescently labeled macromolecular soluble ligands indicated that the LSEC fraction contained 62 x 107 (sd 12 x 107) purified LSEC.Cultured LSEC avidly endocytosed ligands for the mannose and scavenger receptors.CONCLUSIONS: We show here for the first time that pig LSEC, similar to what has been found earlier in rat LSEC, represent an effective scavenger system for removal of macromolecular waste products from the circulation.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Digestive Surgery, University Hospital of Tromsø, 9038 Tromsø, Norway. Geir.Ivar.Nedredal@fagmed.uit.no

ABSTRACT
BACKGROUND: Numerous studies in rats and a few other mammalian species, including man, have shown that the sinusoidal cells constitute an important part of liver function. In the pig, however, which is frequently used in studies on liver transplantation and liver failure models, our knowledge about the function of hepatic sinusoidal cells is scarce. We have explored the scavenger function of pig liver sinusoidal endothelial cells (LSEC), a cell type that in other mammals performs vital elimination of an array of waste macromolecules from the circulation. RESULTS: 125I-macromolecules known to be cleared in the rat via the scavenger and mannose receptors were rapidly removed from the pig circulation, 50% of the injected dose being removed within the first 2-5 min following injection. Fluorescently labeled microbeads (2 &mgr;m in diameter) used to probe phagocytosis accumulated in Kupffer cells only, whereas fluorescently labeled soluble macromolecular ligands for the mannose and scavenger receptors were sequestered only by LSEC. Desmin-positive stellate cells accumulated no probes. Isolation of liver cells using collagenase perfusion through the portal vein, followed by various centrifugation protocols to separate the different liver cell populations yielded 280 x 107 (range 50-890 x 107) sinusoidal cells per liver (weight of liver 237.1 g (sd 43.6)). Use of specific anti-Kupffer cell- and anti-desmin antibodies, combined with endocytosis of fluorescently labeled macromolecular soluble ligands indicated that the LSEC fraction contained 62 x 107 (sd 12 x 107) purified LSEC. Cultured LSEC avidly endocytosed ligands for the mannose and scavenger receptors. CONCLUSIONS: We show here for the first time that pig LSEC, similar to what has been found earlier in rat LSEC, represent an effective scavenger system for removal of macromolecular waste products from the circulation.

No MeSH data available.


Related in: MedlinePlus