scholarly journals Fractionation of Hepatic Nonparenchymal Cells

2002 ◽  
Vol 2 ◽  
pp. 1347-1350 ◽  
Author(s):  
John Graham
Keyword(s):  
Endothelial Cells ◽  
Stellate Cells ◽  
Cell Types ◽  
Low Density ◽  
Nonparenchymal Cells ◽  
Mammalian Liver ◽  
Hepatic Nonparenchymal Cells ◽  
Density Barrier ◽  
Parenchymal Cells ◽  
Different Cell Types

The majority of parenchymal cells from mammalian liver cells can be removed by very low speed centrifugation (50 g) but a simple low-density barrier (1.096 g/ml) is required to remove the remaining parenchymal cells from the 50-g supernatant which contains all of the lower density nonparenchymal cells. Continuous gradients of Nycodenz®can provide satisfactory resolution of Kupffer, stellate, and endothelial cells on an analytical basis but the separation of different cell types is not sufficient preparatively. Flotation through a low-density iodixanol barrier can, however, provide a satisfactory enrichment of the least dense nonparenchymal cell – the stellate cells.

Cytokeratin expression, fibrillar organization, and subtle function in liver cells

1995 ◽  
Vol 73 (9-10) ◽  
pp. 619-625 ◽  
Author(s):  
Normand Marceau ◽  
Anne Loranger
Keyword(s):  
Epithelial Cells ◽  
Cell Types ◽  
Vital Role ◽  
Hepatic Tissue ◽  
Tight Coupling ◽  
Nonparenchymal Cells ◽  
Experimental Approaches ◽  
And Function ◽  
Parenchymal Cells ◽  
Different Cell Types

Cytokeratins (CKs) constitute a diverse group of intermediate filament (IF) proteins, expressed as pairs in keratinized and nonkeratinizing epithelial cells. Much is known now about the expression, assembly, and function of CKs in keratinized epithelial cells, the main features being the tight coupling between CK pair switch and cell terminal differentiation (protection barrier) and the vital role of CK IFs in cell mechanical integrity. However, the picture about nonkeratinizing epithelia, like the hepatic tissue, remains quite unclear. The liver forms a multicellular system, where parenchymal cells (i.e., hepatocytes) exert diverse metabolic function(s) and nonparenchymal epithelial cells (e.g., biliary epithelial cells) usually serve structural (or accessory) purposes. In terms of differential CK gene expression, the data accumulated so far demonstrated that parenchymal cells can contain as few as one single CK pair, whereas nonparenchymal cells contain more than two CKs, one of them being a representative of those found in epidermis. Moreover, the distribution of the CK IF networks present in the different cell types varies a lot and can often be linked to the cell specialization. However, the function(s) played by these IF proteins in this multicellular tissue remains a major issue. The use of new experimental approaches, largely based on gene transfer technology, indicates that it is quite subtle.Key words: cytokeratins, liver, expression, organization, function(s).

scholarly journals A cell based assay for evaluating binding and uptake of an antibody using hepatic nonparenchymal cells

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Yuki Noguchi ◽  
Kazuhisa Ozeki ◽  
Hiroaki Takesue ◽  
Hidetaka Akita
Keyword(s):  
Flow Cytometry ◽  
Endothelial Cells ◽  
Liver Sinusoidal Endothelial Cells ◽  
Nonparenchymal Cells ◽  
A Cell ◽  
Hepatic Nonparenchymal Cells ◽  
Titration Assay ◽  
Parenchymal Cells

AbstractEvaluation of the binding and uptake of an antibody in liver non-parenchymal cells (NPC), including liver sinusoidal endothelial cells, is important for revealing its pharmacokinetic (PK) behavior, since NPC has important roles in eliminating an antibody from the blood via the Fc fragment of IgG receptor IIB (FcγRIIB). However, there is currently no in vitro quantitative assay using NPC. This study reports on the development of a cell-based assay for evaluating the binding and uptake of such an antibody using liver NPC of mice and monkeys. In mice, the FcγRIIB-expressing cells were identified in the CD146-positive and CD45-negative fraction by flow cytometry. A titration assay was performed to determine the PK parameters, and the obtained parameter was comparable to that determined by the fitting of the in vivo PK. This approach was also extended to NPC from monkeys. The concentration-dependent binding and uptake was measured to determine the PK parameters using monkey NPC, the FcγRIIB-expressing fraction of which was identified by CD31 and CD45. The findings presented herein demonstrate that the in vitro liver NPC assay using flow cytometry is a useful tool to determine the binding and uptake of biologics and to predict the PK.

scholarly journals The distribution, induction and isoenzyme profile of glutathione S-transferase and glutathione peroxidase in isolated rat liver parenchymal, Kupffer and endothelial cells

1989 ◽  
Vol 264 (3) ◽  
pp. 737-744 ◽  
Author(s):  
P Steinberg ◽  
H Schramm ◽  
L Schladt ◽  
L W Robertson ◽  
H Thomas ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Glutathione Peroxidase ◽  
Rat Liver ◽  
Peroxidase Activity ◽  
Cell Types ◽  
Transferase Activity ◽  
Glutathione Peroxidase Activity ◽  
Glutathione S Transferase ◽  
Liver Parenchymal ◽  
Parenchymal Cells

The distribution and inducibility of cytosolic glutathione S-transferase (EC 2.5.1.18) and glutathione peroxidase (EC 1.11.1.19) activities in rat liver parenchymal, Kupffer and endothelial cells were studied. In untreated rats glutathione S-transferase activity with 1-chloro-2,4-dinitrobenzene and 4-hydroxynon-2-trans-enal as substrates was 1.7-2.2-fold higher in parenchymal cells than in Kupffer and endothelial cells, whereas total, selenium-dependent and non-selenium-dependent glutathione peroxidase activities were similar in all three cell types. Glutathione S-transferase isoenzymes in parenchymal and non-parenchymal cells isolated from untreated rats were separated by chromatofocusing in an f.p.l.c. system: all glutathione S-transferase isoenzymes observed in the sinusoidal lining cells were also detected in the parenchymal cells, whereas Kupffer and endothelial cells lacked several glutathione S-transferase isoenzymes present in parenchymal cells. At 5 days after administration of Arocolor 1254 glutathione S-transferase activity was only enhanced in parenchymal cells; furthermore, selenium-dependent glutathione peroxidase activity decreased in parenchymal and non-parenchymal cells. At 13 days after a single injection of Aroclor 1254 a strong induction of glutathione S-transferase had taken place in all three cell types, whereas selenium-dependent glutathione peroxidase activity remained unchanged (endothelial cells) or was depressed (parenchymal and Kupffer cells). Hence these results clearly establish that glutathione S-transferase and glutathione peroxidase are differentially regulated in rat liver parenchymal as well as non-parenchymal cells. The presence of glutathione peroxidase and several glutathione S-transferase isoenzymes capable of detoxifying a variety of compounds in Kupffer and endothelial cells might be crucial to protect the liver from damage by potentially hepatotoxic substances.

EXPRESSION OF PLATELET ALLOANTIGENS ON HUMAN ENDOTHELIAL CELLS AND HEL CELLS

1987 ◽  
Author(s):  
Y Kawai ◽  
R R Montgomery ◽  
K Furihata ◽  
T J Kunicki
Keyword(s):  
Endothelial Cells ◽  
Protein A ◽  
Cell Types ◽  
Membrane Glycoproteins ◽  
Human Endothelial Cells ◽  
Immunoblot Assay ◽  
Sds Page ◽  
Hel Cells ◽  
Erythroleukemia Cell ◽  
Different Cell Types

Analogs of platelet membrane glycoproteins IIb and IIIa (GPIIb-IIIa) have been shown to be synthesized and expressed by human endothelial cells (HEC), a human erythroleukemia cell line (HEL) and various other cells. Previous studies from our laboratory demonstrated that the platelet alloantigen P1A1, is expressed on HEC GPIIIa. Other alloantigen systems, namely, Pen and Bak, are known to be localized on platelet GPIIIa and GPIIb, respectively. Utilizing additional antibodies from patients with PTP specific for Pena, Baka, and Bakb allo-antigens, and isoantibodies (iso-ab) from a patient with Glanzmann's Thrombasthenia (GT), we have studied cultured HEC and HEL cells for expression of epitopes recognized by these antibodies. HEC and HEL cells were meta-bolically labeled with 35S-methionine and lysed in 0.5% TX-100 in the presence of 5mM EDTA. Soluble antigens were immunoprecipitated with these antibodies coupled to Protein A-Sepharose and subjected to SDS-PAGE and fluorography. Anti-Pena and the GT iso-ab reacted with the GPIIb-IIIa complex from both HEC and HEL lysates, but anti-Baka and anti-Bakb failed to immunoprecipitate GPIIb-IIIa analogs from either HEC or HEL. In an immunoblot assay, the GT iso-ab bound to GPIIIa of both HEC and HEL. Anti-Pena failed to react with SDS-denatured proteins. HEL GPIIIa migrates slightly faster than HEC GPIIIa and slightly slower than platelet GPIIIa. These results indicate that the epitopes of platelet GPIIIa recognized by alloantibodies and isoantibodies are shared by GPIIIa analogs of HEC and HEL. GPIIb-associated alloantigens are not expressed by HEC and HEL GPIIb analogs, an observation that is consistent with the decreased structural homology between GPIIb analogs derived from different cell types.

Rat liver endothelial cell glutamine transporter and glutaminase expression contrast with parenchymal cells

1999 ◽  
Vol 276 (3) ◽  
pp. G743-G750 ◽  
Author(s):  
Rüdiger Lohmann ◽  
Wiley W. Souba ◽  
Barrie P. Bode
Keyword(s):  
Endothelial Cells ◽  
Amino Acid ◽  
Cell Types ◽  
Male Rats ◽  
Specific Cell ◽  
Liver Endothelial Cell ◽  
Glutamine Transporter ◽  
Health And Disease ◽  
Parenchymal Cells ◽  
Functional Analyses

Despite the central role of the liver in glutamine homeostasis in health and disease, little is known about the mechanism by which this amino acid is transported into sinusoidal endothelial cells, the second most abundant hepatic cell type. To address this issue, the transport ofl-glutamine was functionally characterized in hepatic endothelial cells isolated from male rats. On the basis of functional analyses, including kinetics, cation substitution, and amino acid inhibition, it was determined that a Na+-dependent carrier distinct from system N in parenchymal cells, with properties of system ASC or B0, mediated the majority of glutamine transport in hepatic endothelial cells. These results were supported by Northern blot analyses that showed expression of the ATB0 transporter gene in endothelial but not parenchymal cells. Concurrently, it was determined that, whereas both cell types express glutamine synthetase, hepatic endothelial cells express the kidney-type glutaminase isozyme in contrast to the liver-type isozyme in parenchymal cells. This represents the first report of ATB0 and kidney-type glutaminase isozyme expression in the liver, observations that have implications for roles of specific cell types in hepatic glutamine homeostasis in health and disease.

scholarly journals Multi-cellular 3D human primary liver cell culture elevates metabolic activity under fluidic flow

Lab on a Chip ◽  
2015 ◽  
Vol 15 (10) ◽  
pp. 2269-2277 ◽  
Author(s):  
Mandy B. Esch ◽  
Jean-Matthieu Prot ◽  
Ying I. Wang ◽  
Paula Miller ◽  
Jose Ricardo Llamas-Vidales ◽  
...  
Keyword(s):  
Cell Culture ◽  
Metabolic Activity ◽  
Kupffer Cells ◽  
Liver Cell ◽  
Low Cost ◽  
Stellate Cells ◽  
Cell Types ◽  
Liver Cell Culture ◽  
Parenchymal Cells

We have developed a low-cost liver cell culture device that creates fluidic flow over a 3D primary liver cell culture that consists of multiple liver cell types, including hepatocytes and non-parenchymal cells (fibroblasts, stellate cells, and Kupffer cells).

Cell-derived microparticles in haemostasis and vascular medicine

2009 ◽  
Vol 101 (03) ◽  
pp. 439-451 ◽  
Author(s):  
Laurent Burnier ◽  
Pierre Fontana ◽  
Brenda R. Kwak ◽  
Anne Angelillo-Scherrer
Keyword(s):  
Endothelial Cells ◽  
Red Blood Cells ◽  
Blood Cells ◽  
Essential Role ◽  
Cell Types ◽  
Vascular Medicine ◽  
Disease Markers ◽  
Wide Range ◽  
Definition Of ◽  
Different Cell Types

SummaryConsiderable interest for cell-derived microparticles has emerged, pointing out their essential role in haemostatic response and their potential as disease markers, but also their implication in a wide range of physiological and pathological processes. They derive from different cell types including platelets – the main source of microparticles – but also from red blood cells, leukocytes and endothelial cells, and they circulate in blood. Despite difficulties encountered in analyzing them and disparities of results obtained with a wide range of methods, microparticle generation processes are now better understood. However, a generally admitted definition of microparticles is currently lacking. For all these reasons we decided to review the literature regarding microparticles in their widest definition, including ectosomes and exosomes, and to focus mainly on their role in haemostasis and vascular medicine.

The Application of Quantitative Stereology to the Study of Adenohypophyseal Cells

1976 ◽  
Vol 34 ◽  
pp. 124-125
Author(s):  
Max C. Poole ◽  
V.B. Mahesh ◽  
Allen Costoff
Keyword(s):  
Anterior Pituitary ◽  
Cell Types ◽  
The Other ◽  
Vaginal Smears ◽  
Prolactin Cells ◽  
Estrous Cycles ◽  
Liver Parenchymal ◽  
Quantitative Stereology ◽  
Parenchymal Cells ◽  
Different Cell Types

Quantitative stereology of liver parenchymal cells has previously been reported (1,2), but there have been few studies of morphometry applied to a heterogenous tissue (3). Due to the presence of several different cell types, it is difficult to study the synthesis and secretion of hormones in cells of the anterior pituitary by conventional biochemical means. In this study prolactin cells were analyzed using morphometry during different times of the rat estrous cycle, and were correlated with changing levels of prolactin in the serum and pituitary gland.Vaginal smears of 60 day old Holtzman rats were monitored through three estrous cycles, and only four day cycling rats were used. Groups of six animals were decapitated at 4 P.M., 6 P.M., 10 P.M. and 12 midnight of proestrus and one half of the pituitary was processed for electron microscopy and the other half for assay.

scholarly journals Endocytosis of ricin by rat liver cells in vivo and in vitro is mainly mediated by mannose receptors on sinusoidal endothelial cells

1993 ◽  
Vol 291 (3) ◽  
pp. 749-755 ◽  
Author(s):  
S Magnússon ◽  
T Berg
Keyword(s):  
Endothelial Cells ◽  
Mannose Receptor ◽  
Cell Types ◽  
Plant Toxin ◽  
Sinusoidal Endothelial Cells ◽  
Liver Uptake ◽  
Parenchymal Cells ◽  
Mannose Receptors

Upon intravenous injection into rats, the plant toxin ricin was rapidly cleared from the circulation by the liver. Among the different liver cell populations, most of the injected ricin associated with the sinusoidal endothelial cells (EC), whereas the liver parenchymal cells (PC) and Kupffer cells (KC) yielded minor contributions to the total liver uptake in vivo. Co-injection of mannan strongly inhibited ricin uptake by the EC, showing that it was mediated by mannose receptors. On the other hand, co-injection of lactose, which inhibits the galactose-specific association of ricin with cells, enhanced ricin uptake by the EC. The carbohydrate-dependency of the EC contribution to the uptake of ricin in vivo was reflected in the carbohydrate-dependency of the uptake in vivo by whole liver. In vitro, the EC also endocytosed ricin more efficiently than did the PC or KC. Whereas uptake in vitro in the EC was mainly mannose-specific, uptake in the two other cell types was mainly galactose-specific. Western blotting showed that the mannose receptors of liver non-parenchymal cells are identical with the mannose receptor previously isolated from alveolar macrophages. The mannose receptors are expressed at a higher level in EC than in KC. Ligand blotting showed that, in the presence of lactose, the mannose receptor is the only protein in the EC that binds ricin, and the binding is mannose-specific and Ca(2+)-dependent.

scholarly journals The uptake and metabolism of chylomicron-remnant lipids by rat liver parenchymal and non-parenchymal cells in vitro

1985 ◽  
Vol 232 (2) ◽  
pp. 395-401 ◽  
Author(s):  
P M Lippiello ◽  
P J Sisson ◽  
M Waite
Keyword(s):  
Endothelial Cells ◽  
Rat Liver ◽  
Kupffer Cells ◽  
Liver Cell ◽  
Cholesterol Ester ◽  
Cell Types ◽  
Chylomicron Remnant ◽  
Parenchymal Cells ◽  
Uptake And Metabolism

The uptake and metabolism of chylomicron-remnant lipids by individual liver cell types was examined by incubating remnants with monolayer cultures of hepatocytes, Kupffer cells, and endothelial cells from rat liver. Remnants were prepared in vitro from radiolabelled mesenteric-lymph chylomicra, utilizing either purified lipoprotein lipase from bovine milk, or plasma isolated from heparinized rats. The resulting particles contained [3H]phosphatidylcholine and cholesterol, and [14C]oleate in the acylglycerol, phospholipid, fatty-acid and cholesterol-ester fractions. The capacities of the three cell types for uptake of both [3H]lipids and [14C]lipids were determined to be, on a per-cell basis, in the order: Kupffer greater than hepatocytes greater than endothelial. The relative proportions of [3H]phospholipid and total [3H]cholesterol taken up by hepatocytes and non-parenchymal cells remained constant with time. The uptake of [14C]oleoyl lipids by all three cell types was slightly greater than that of the total [3H]cholesterol and [3H]phospholipid components. There was evidence of cholesterol-ester hydrolysis and turnover of [14C]oleate in the phospholipid fraction in hepatocytes and Kupffer cells, but not endothelial cells, over the first 2 h. With both remnant preparations, these observations indicate that significant differences exist between the three major liver cell types with respect to the uptake and metabolism of remnant lipid components.

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