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Article ; Online: Old Drugs to Reinvigorate Old Liver Cells.

Smedsrød, Bård

The journals of gerontology. Series A, Biological sciences and medical sciences

2020 Volume 75, Issue 2, Page(s) 266–267

MeSH term(s)	Aged ; Humans ; Liver ; Reperfusion Injury/drug therapy
Language	English
Publishing date	2020-01-03
Publishing country	United States
Document type	Journal Article ; Comment
ZDB-ID	1223643-3
ISSN	1758-535X ; 1079-5006
ISSN (online)	1758-535X
ISSN	1079-5006
DOI	10.1093/gerona/glz278
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Article ; Online: Protocol for Isolation and Culture of Mouse Hepatocytes (HCs), Kupffer Cells (KCs), and Liver Sinusoidal Endothelial Cells (LSECs) in Analyses of Hepatic Drug Distribution.

Elvevold, Kjetil / Kyrrestad, Ingelin / Smedsrød, Bård

Methods in molecular biology (Clifton, N.J.)

2022 Volume 2434, Page(s) 385–402

Abstract: Development of the new generation of drugs (e.g., oligo- and polynucleotides administered intravascularly either as free compounds or as nano-formulations) frequently encounters major challenges such as lack of control of targeting and/or delivery. ... ...

Abstract	Development of the new generation of drugs (e.g., oligo- and polynucleotides administered intravascularly either as free compounds or as nano-formulations) frequently encounters major challenges such as lack of control of targeting and/or delivery. Uncontrolled or unwanted clearance by the liver is a well-known and particularly important hurdle in this respect. Hence, reliable techniques are needed to identify the type(s) of liver cells, receptors, and metabolic mechanisms that are responsible for unwanted clearance of these compounds.We describe here a method for the isolation and culture of the major cell types from mouse liver: hepatocytes (HCs), Kupffer cells (KCs), and liver sinusoidal endothelial cells (LSECs). The presently described protocol employs perfusion of the liver with a collagenase-based enzyme preparation to effectively transform the intact liver to a single cell suspension. From this initial cell suspension HCs are isolated by specified centrifugation schemes, yielding highly pure HC preparations, and KCs and LSECs are isolated by employing magnetic-activated cell sorting (MACS). The MACS protocol makes use of magnetic microbeads conjugated with specific antibodies that bind unique surface antigens on either KCs or LSECs. In this way the two cell types are specifically and separately pulled out of the initial liver cell suspension by applying a magnetic field, resulting in high purity, yield, and viability of the two cell types, allowing functional studies of the cells.If the drug compound in question is to be studied with respect to liver cell distribution of intravascularly administered drug compounds the isolated cells can be analyzed directly after isolation. Detailed studies of receptor-ligand interactions and/or dynamics of intracellular metabolism of the compound can be conducted in primary surface cultures of HCs, LSECs, and KCs established by seeding the isolated cells on specified growth substrates.
MeSH term(s)	Animals ; Endothelial Cells/metabolism ; Hepatocytes/metabolism ; Kupffer Cells ; Liver/metabolism ; Mice ; Pharmaceutical Preparations/metabolism
Chemical Substances	Pharmaceutical Preparations
Language	English
Publishing date	2022-02-25
Publishing country	United States
Document type	Journal Article
ISSN	1940-6029
ISSN (online)	1940-6029
DOI	10.1007/978-1-0716-2010-6_27
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Article ; Online: Liver sinusoidal cells eliminate blood-borne phage K1F.

Sánchez Romano, Javier / Simón-Santamaría, Jaione / McCourt, Peter / Smedsrød, Bård / Mortensen, Kim Erlend / Sagona, Antonia P / Sørensen, Karen Kristine / Larsen, Anett Kristin

mSphere

2024 Volume 9, Issue 3, Page(s) e0070223

Abstract: Phage treatment has regained attention due to an increase in multiresistant bacteria. For phage therapy to be successful, phages must reach their target bacteria in sufficiently high numbers. Blood-borne phages are believed to be captured by macrophages ... ...

Abstract	Phage treatment has regained attention due to an increase in multiresistant bacteria. For phage therapy to be successful, phages must reach their target bacteria in sufficiently high numbers. Blood-borne phages are believed to be captured by macrophages in the liver and spleen. Since liver sinusoids also consist of specialized scavenger liver sinusoidal endothelial cells (LSECs) and Kupffer cells (KCs), this study investigated the contribution of both cell types in the elimination of
MeSH term(s)	Mice ; Humans ; Animals ; Bacteriophages ; Endothelial Cells ; Hepatocytes ; Liver ; Endocytosis ; Mammals
Language	English
Publishing date	2024-02-28
Publishing country	United States
Document type	Journal Article
ISSN	2379-5042
ISSN (online)	2379-5042
DOI	10.1128/msphere.00702-23
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Article: The Scavenger Function of Liver Sinusoidal Endothelial Cells in Health and Disease.

Bhandari, Sabin / Larsen, Anett Kristin / McCourt, Peter / Smedsrød, Bård / Sørensen, Karen Kristine

Frontiers in physiology

2021 Volume 12, Page(s) 757469

Abstract: The aim of this review is to give an outline of the blood clearance function of the liver sinusoidal endothelial cells (LSECs) in health and disease. Lining the hundreds of millions of hepatic sinusoids in the human liver the LSECs are perfectly located ... ...

Abstract	The aim of this review is to give an outline of the blood clearance function of the liver sinusoidal endothelial cells (LSECs) in health and disease. Lining the hundreds of millions of hepatic sinusoids in the human liver the LSECs are perfectly located to survey the constituents of the blood. These cells are equipped with high-affinity receptors and an intracellular vesicle transport apparatus, enabling a remarkably efficient machinery for removal of large molecules and nanoparticles from the blood, thus contributing importantly to maintain blood and tissue homeostasis. We describe here central aspects of LSEC signature receptors that enable the cells to recognize and internalize blood-borne waste macromolecules at great speed and high capacity. Notably, this blood clearance system is a silent process, in the sense that it usually neither requires or elicits cell activation or immune responses. Most of our knowledge about LSECs arises from studies in animals, of which mouse and rat make up the great majority, and some species differences relevant for extrapolating from animal models to human are discussed. In the last part of the review, we discuss comparative aspects of the LSEC scavenger functions and specialized scavenger endothelial cells (SECs) in other vascular beds and in different vertebrate classes. In conclusion, the activity of LSECs and other SECs prevent exposure of a great number of waste products to the immune system, and molecules with noxious biological activities are effectively "silenced" by the rapid clearance in LSECs. An undesired consequence of this avid scavenging system is unwanted uptake of nanomedicines and biologics in the cells. As the development of this new generation of therapeutics evolves, there will be a sharp increase in the need to understand the clearance function of LSECs in health and disease. There is still a significant knowledge gap in how the LSEC clearance function is affected in liver disease.
Language	English
Publishing date	2021-10-11
Publishing country	Switzerland
Document type	Journal Article ; Review
ZDB-ID	2564217-0
ISSN	1664-042X
ISSN	1664-042X
DOI	10.3389/fphys.2021.757469
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Article ; Online: Changes in the proteome and secretome of rat liver sinusoidal endothelial cells during early primary culture and effects of dexamethasone.

Li, Ruomei / Bhandari, Sabin / Martinez-Zubiaurre, Inigo / Bruun, Jack-Ansgar / Urbarova, Ilona / Smedsrød, Bård / Simón-Santamaría, Jaione / Sørensen, Karen Kristine

PloS one

2022 Volume 17, Issue 9, Page(s) e0273843

Abstract: Introduction: Liver sinusoidal endothelial cells (LSECs) are specialized fenestrated scavenger endothelial cells involved in the elimination of modified plasma proteins and tissue turnover waste macromolecules from blood. LSECs also participate in liver ...

Abstract	Introduction: Liver sinusoidal endothelial cells (LSECs) are specialized fenestrated scavenger endothelial cells involved in the elimination of modified plasma proteins and tissue turnover waste macromolecules from blood. LSECs also participate in liver immune responses. A challenge when studying LSEC biology is the rapid loss of the in vivo phenotype in culture. In this study, we have examined biological processes and pathways affected during early-stage primary culture of rat LSECs and checked for cell responses to the pro-inflammatory cytokine interleukin (IL)-1β and the anti-inflammatory drug dexamethasone. Methods: LSECs from male Sprague Dawley rats were cultured on type I collagen in 5% oxygen atmosphere in DMEM with serum-free supplements for 2 and 24 h. Quantitative proteomics using tandem mass tag technology was used to examine proteins in cells and supernatants. Validation was done with qPCR, ELISA, multiplex immunoassay, and caspase 3/7 assay. Cell ultrastructure was examined by scanning electron microscopy, and scavenger function by quantitative endocytosis assays. Results: LSECs cultured for 24 h showed a characteristic pro-inflammatory phenotype both in the presence and absence of IL-1β, with upregulation of cellular responses to cytokines and interferon-γ, cell-cell adhesion, and glycolysis, increased expression of fatty acid binding proteins (FABP4, FABP5), and downregulation of several membrane receptors (STAB1, STAB2, LYVE1, CLEC4G) and proteins in pyruvate metabolism, citric acid cycle, fatty acid elongation, amino acid metabolism, and oxidation-reduction processes. Dexamethasone inhibited apoptosis and improved LSEC viability in culture, repressed inflammatory and immune regulatory pathways and secretion of IL-1β and IL-6, and further upregulated FABP4 and FABP5 compared to time-matched controls. The LSEC porosity and endocytic activity were reduced at 24 h both with and without dexamethasone but the dexamethasone-treated cells showed a less stressed phenotype. Conclusion: Rat LSECs become activated towards a pro-inflammatory phenotype during early culture. Dexamethasone represses LSEC activation, inhibits apoptosis, and improves cell viability.
MeSH term(s)	Animals ; Dexamethasone/metabolism ; Dexamethasone/pharmacology ; Endothelial Cells/metabolism ; Liver/metabolism ; Male ; Proteome/metabolism ; Rats ; Rats, Sprague-Dawley ; Secretome
Chemical Substances	Proteome ; Dexamethasone (7S5I7G3JQL)
Language	English
Publishing date	2022-09-02
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2267670-3
ISSN	1932-6203 ; 1932-6203
ISSN (online)	1932-6203
ISSN	1932-6203
DOI	10.1371/journal.pone.0273843
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Article ; Online: Liver sinusoidal endothelial cells show reduced scavenger function and downregulation of Fc gamma receptor IIb, yet maintain a preserved fenestration in the Glmpgt/gt mouse model of slowly progressing liver fibrosis.

Antwi, Milton Boaheng / Dumitriu, Gianina / Simón-Santamaria, Jaione / Romano, Javier Sánchez / Li, Ruomei / Smedsrød, Bård / Vik, Anders / Eskild, Winnie / Sørensen, Karen Kristine

PloS one

2023 Volume 18, Issue 11, Page(s) e0293526

Abstract: Liver sinusoidal endothelial cells (LSECs) are fenestrated endothelial cells with a unique, high endocytic clearance capacity for blood-borne waste macromolecules and colloids. This LSEC scavenger function has been insufficiently characterized in liver ... ...

Abstract	Liver sinusoidal endothelial cells (LSECs) are fenestrated endothelial cells with a unique, high endocytic clearance capacity for blood-borne waste macromolecules and colloids. This LSEC scavenger function has been insufficiently characterized in liver disease. The Glmpgt/gt mouse lacks expression of a subunit of the MFSD1/GLMP lysosomal membrane protein transporter complex, is born normal, but soon develops chronic, mild hepatocyte injury, leading to slowly progressing periportal liver fibrosis, and splenomegaly. This study examined how LSEC scavenger function and morphology are affected in the Glmpgt/gt model. FITC-labelled formaldehyde-treated serum albumin (FITC-FSA), a model ligand for LSEC scavenger receptors was administered intravenously into Glmpgt/gt mice, aged 4 months (peak of liver inflammation), 9-10 month, and age-matched Glmpwt/wt mice. Organs were harvested for light and electron microscopy, quantitative image analysis of ligand uptake, collagen accumulation, LSEC ultrastructure, and endocytosis receptor expression (also examined by qPCR and western blot). In both age groups, the Glmpgt/gt mice showed multifocal liver injury and fibrosis. The uptake of FITC-FSA in LSECs was significantly reduced in Glmpgt/gt compared to wild-type mice. Expression of LSEC receptors stabilin-1 (Stab1), and mannose receptor (Mcr1) was almost similar in liver of Glmpgt/gt mice and age-matched controls. At the same time, immunostaining revealed differences in the stabilin-1 expression pattern in sinusoids and accumulation of stabilin-1-positive macrophages in Glmpgt/gt liver. FcγRIIb (Fcgr2b), which mediates LSEC endocytosis of soluble immune complexes was widely and significantly downregulated in Glmpgt/gt liver. Despite increased collagen in space of Disse, LSECs of Glmpgt/gt mice showed well-preserved fenestrae organized in sieve plates but the frequency of holes >400 nm in diameter was increased, especially in areas with hepatocyte damage. In both genotypes, FITC-FSA also distributed to endothelial cells of spleen and bone marrow sinusoids, suggesting that these locations may function as possible compensatory sites of clearance of blood-borne scavenger receptor ligands in liver fibrosis.
MeSH term(s)	Mice ; Animals ; Endothelial Cells/metabolism ; Ligands ; Down-Regulation ; Fluorescein-5-isothiocyanate/metabolism ; Liver/metabolism ; Liver Cirrhosis/genetics ; Liver Cirrhosis/metabolism ; Hepatocytes/metabolism ; Disease Models, Animal ; Collagen/metabolism ; Membrane Transport Proteins/metabolism ; Cell Adhesion Molecules, Neuronal/genetics ; Cell Adhesion Molecules, Neuronal/metabolism
Chemical Substances	Fc gamma receptor IIB ; Ligands ; Fluorescein-5-isothiocyanate (I223NX31W9) ; Collagen (9007-34-5) ; MFSD1 protein, mouse ; Membrane Transport Proteins ; Stab1 protein, mouse ; Cell Adhesion Molecules, Neuronal
Language	English
Publishing date	2023-11-01
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2267670-3
ISSN	1932-6203 ; 1932-6203
ISSN (online)	1932-6203
ISSN	1932-6203
DOI	10.1371/journal.pone.0293526
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Article: Clearance function of scavenger endothelial cells.

Smedsrød, Bård

Comparative hepatology

2004 Volume 3 Suppl 1, Page(s) S22

Language	English
Publishing date	2004-01-14
Publishing country	England
Document type	Journal Article
ISSN	1476-5926
ISSN	1476-5926
DOI	10.1186/1476-5926-2-S1-S22
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Article ; Online: Evaluating liver uptake and distribution of different poly(2-methyl-2-oxazoline) modified lysine dendrimers following intravenous administration.

England, Richard M / Moss, Jennifer I / Hill, Kathryn J / Elvevold, Kjetil / Smedsrød, Bård / Ashford, Marianne B

Biomaterials science

2019 Volume 7, Issue 8, Page(s) 3418–3424

Abstract: We report on the synthesis of four poly(2-methyl-2-oxazoline) modified lysine dendrimers with different residual groups or modifications on the dendrimer core, including: amino groups (positive charge), carboxyl groups (negative charge), and two drug ... ...

Abstract	We report on the synthesis of four poly(2-methyl-2-oxazoline) modified lysine dendrimers with different residual groups or modifications on the dendrimer core, including: amino groups (positive charge), carboxyl groups (negative charge), and two drug molecules, one of which has a high log P. We looked at the in vivo distribution amongst three main liver cell types: hepatocytes, liver sinusoidal endothelial cells (LSECs) and Kupffer cells (KCs) and found differences in cell distribution and uptake concentrations dependent on these residual groups. In particular, the amino-functional polymer showed greater uptake by the hepatocytes whilst the carboxyl-functionalised polymer exhibited greater uptake by KCs and LSECs. These findings provide insight into which professional scavenger cells of the liver remove these types of nanoparticles from the bloodstream and we describe some of the design criteria to consider when creating novel drug delivery systems.
MeSH term(s)	Administration, Intravenous ; Animals ; Biological Transport ; Dendrimers/chemistry ; Female ; Hydrophobic and Hydrophilic Interactions ; Liver/metabolism ; Lysine/administration & dosage ; Lysine/chemistry ; Lysine/metabolism ; Lysine/pharmacokinetics ; Mice ; Polyamines/chemistry ; Rhodamines/chemistry ; Tissue Distribution
Chemical Substances	Dendrimers ; Polyamines ; Rhodamines ; poly(2-methyl-2-oxazoline) ; Lysine (K3Z4F929H6)
Language	English
Publishing date	2019-07-03
Publishing country	England
Document type	Journal Article
ZDB-ID	2693928-9
ISSN	2047-4849 ; 2047-4830
ISSN (online)	2047-4849
ISSN	2047-4830
DOI	10.1039/c9bm00385a
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Article ; Online: Autofluorescence in freshly isolated adult human liver sinusoidal cells.

Larsen, Anett Kristin / Simón-Santamaría, Jaione / Elvevold, Kjetil / Ericzon, Bo Göran / Mortensen, Kim Erlend / McCourt, Peter / Smedsrød, Bård / Sørensen, Karen Kristine

European journal of histochemistry : EJH

2021 Volume 65, Issue 4

Abstract: Autofluorescent granules of various sizes were observed in primary human liver endothelial cells (LSECs) upon laser irradiation using a wide range of wavelengths. Autofluorescence was detected in LAMP-1 positive vesicles, suggesting lysosomal location. ... ...

Abstract	Autofluorescent granules of various sizes were observed in primary human liver endothelial cells (LSECs) upon laser irradiation using a wide range of wavelengths. Autofluorescence was detected in LAMP-1 positive vesicles, suggesting lysosomal location. Confocal imaging of freshly prepared cultures and imaging flow cytometry of non-cultured cells revealed fluorescence in all channels used. Treatment with a lipofuscin autofluorescence quencher reduced autofluorescence, most efficiently in the near UV-area. These results, combined with the knowledge of the very active blood clearance function of LSECs support the notion that lysosomally located autofluorescent material reflected accumulation of lipofuscin in the intact liver. These results illustrate the importance of careful selection of fluorophores, especially when labelling of live cells where the quencher is not compatible.
MeSH term(s)	Adult ; Endothelial Cells/cytology ; Endothelial Cells/metabolism ; Fluorescence ; Humans ; Lipofuscin/metabolism ; Liver/cytology ; Liver/metabolism ; Microscopy, Fluorescence
Chemical Substances	Lipofuscin
Language	English
Publishing date	2021-12-13
Publishing country	Italy
Document type	Journal Article
ZDB-ID	1109511-8
ISSN	2038-8306 ; 0391-7258 ; 1121-4201 ; 1121-760X
ISSN (online)	2038-8306
ISSN	0391-7258 ; 1121-4201 ; 1121-760X
DOI	10.4081/ejh.2021.3337
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Article ; Online: Liver sinusoidal endothelial cell ICAM-1 mediated tumor/endothelial crosstalk drives the development of liver metastasis by initiating inflammatory and angiogenic responses.

Benedicto, Aitor / Herrero, Alba / Romayor, Irene / Marquez, Joana / Smedsrød, Bård / Olaso, Elvira / Arteta, Beatriz

Scientific reports

2019 Volume 9, Issue 1, Page(s) 13111

Abstract: The prometastatic stroma generated through tumor cells/host cells interaction is critical for metastatic growth. To elucidate the role of ICAM-1 on the crosstalk between tumor and primary liver sinusoidal endothelial cells (LSECs) and hepatic stellate ... ...

Abstract	The prometastatic stroma generated through tumor cells/host cells interaction is critical for metastatic growth. To elucidate the role of ICAM-1 on the crosstalk between tumor and primary liver sinusoidal endothelial cells (LSECs) and hepatic stellate cells (HSCs), implicated in tumor adhesion and angiogenesis, we performed in vitro cocultures and an in vivo model of liver metastasis of colorectal cancer (CRC). ICAM-1 blockade in the LSECs decreased the adhesion and transmigration of tumor cells through an LSEC in vitro and vivo. Cocultures of C26 cells and LSECs contained higher amounts of IL-1β, IL-6, PGE-2, TNF-α and ICAM-1 than monocultures. C26 cells incubated with sICAM-1 secreted higher amounts of PGE-2, IL-6, VEGF, and MMPs, while enhanced the migration of LSECs and HSCs. HSCs cultures activated by media from C26 cells pretreated with sICAM-1 contained the largest amounts of VEGF and MMPs. C26 cell activation with sICAM-1 enhanced their metastasizing potential in vivo, while tumor LFA-1 blockade reduced tumor burden and LSECs and HSC-derived myofibroblasts recruitment. In vivo ICAM-1 silencing produced similar results. These findings uncover LSEC ICAM-1 as a mediator of the CRC metastatic cascade in the liver and identifies it as target for the inhibition of liver colonization and metastatic progression.
MeSH term(s)	Animals ; Apoptosis ; Biomarkers, Tumor/genetics ; Biomarkers, Tumor/metabolism ; Capillaries/immunology ; Capillaries/metabolism ; Capillaries/pathology ; Cell Adhesion ; Cell Communication ; Cell Movement ; Cell Proliferation ; Colonic Neoplasms/etiology ; Colonic Neoplasms/metabolism ; Colonic Neoplasms/pathology ; Endothelial Cells/immunology ; Endothelial Cells/metabolism ; Endothelial Cells/pathology ; Gene Expression Regulation, Neoplastic ; Humans ; Inflammation/complications ; Inflammation/immunology ; Intercellular Adhesion Molecule-1/genetics ; Intercellular Adhesion Molecule-1/metabolism ; Liver Neoplasms/etiology ; Liver Neoplasms/metabolism ; Liver Neoplasms/secondary ; Male ; Mice ; Mice, Inbred BALB C ; Neovascularization, Pathologic/complications ; Neovascularization, Pathologic/immunology ; Tumor Cells, Cultured ; Xenograft Model Antitumor Assays
Chemical Substances	Biomarkers, Tumor ; ICAM1 protein, human ; Intercellular Adhesion Molecule-1 (126547-89-5)
Language	English
Publishing date	2019-09-11
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2615211-3
ISSN	2045-2322 ; 2045-2322
ISSN (online)	2045-2322
ISSN	2045-2322
DOI	10.1038/s41598-019-49473-7
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