scholarly journals Comparison of the ability of basement membranes produced by corneal endothelial and mouse-derived Endodermal PF-HR-9 cells to support the proliferation and differentiation of bovine kidney tubule epithelial cells in vitro.

1984 ◽  
Vol 99 (3) ◽  
pp. 947-961 ◽  
Author(s):  
D Gospodarowicz ◽  
J Lepine ◽  
S Massoglia ◽  
I Wood
Keyword(s):  
Epithelial Cells ◽  
Basement Membrane ◽  
Cell Monolayer ◽  
Basement Membranes ◽  
Low Density ◽  
Kidney Tubule ◽  
Confluent Monolayer ◽  
Bovine Kidney ◽  
Fibroblastic Cells

The proliferation and morphological differentiation of bovine kidney collecting-tubule epithelial cells has been examined as a function of substrata and plasma factors. Collecting kidney tubule explant maintained in vitro gave rise to two distinct cell populations; one was composed mostly of fibroblastic cells whereas the other was epithelioid (EP cells). The proliferation of fibroblastic cells when exposed to serum-supplemented medium was best expressed when cells were maintained on a basement membrane produced by bovine corneal endothelial cells. This basement membrane has a composition, which in previous studies has been shown to favor the proliferation of mesenchymal cells. In contrast, the proliferation of EP cells was best expressed when cells were maintained on a basement membrane produced by the mouse-derived endodermal cell line PF-HR-9 (HR-9-BM). This basement membrane has a biochemical composition very similar to the basement membrane underlying the kidney tubules. Although the fibroblast confluent monolayer maintained on bovine corneal endothelial cell extracellular matrix did not undergo morphogenesis, the confluent monolayer of EP cells maintained on HR-9-BM shows hemicyst formation, suggesting that they were capable of vectorial fluid transport. They also built a complex three-dimensional kidney tubulelike network. Some tubules became grossly visible and floated into the tissue culture medium, remaining tethered to the cell monolayer at either end of the tubule. On an ultrastructural level, the tubules consisted of cells held together with junctional complexes arranged so as to form a lumen. The smallest lumen were bordered by 2-3 cells, and the largest ones by 8-15 cells. The lumens of the larger tubules did contain granular fibrillar and amorphous debris. Low-density EP cell cultures maintained on HR-9-BM could be induced to proliferate at a rate approaching that of cultures exposed to serum when they were exposed to medium supplemented with high-density lipoprotein (HDL, 750 micrograms protein/ml) and transferrin (50 micrograms/ml). When exposed to HDL concentrations equal or lower than 250 micrograms protein/ml, low-density cultures proliferated at a slow rate and readily formed tubulelike structures. This observation indicates that EP cells do not need to reach confluence to undergo morphogenesis, and that HDL, which in the presence of transferrin supports the cell proliferation, can favor their differentiation into tubulelike structures once its concentration becomes limiting for mitogenesis.

scholarly journals Die Kaliumhydrogenphosphat-induzierte Nephropathie des Hundes. I. Pathogenese der Tubulusatrophiee [Potassium Hydrogen Phosphate Induced Nephropathy in the Dog. I. Pathogenesis of Tubular Atrophy—author's trans.]

1980 ◽  
Vol 17 (6) ◽  
pp. 699-719 ◽  
Author(s):  
P. Schneider ◽  
G. Pappritz ◽  
R. Müller-Peddinghaus ◽  
M. Bauer ◽  
H. Lehmann ◽  
...  
Keyword(s):  
Acid Phosphatase ◽  
Epithelial Cells ◽  
Basement Membrane ◽  
Basement Membranes ◽  
Tubular Epithelial Cells ◽  
Beagle Dogs ◽  
Lysosomal Degradation ◽  
Tubular Basement Membrane ◽  
Tubular Atrophy ◽  
Study Results

A nephropathy with severe tubular atrophy was observed in Beagle dogs after oral administration of K2HPO4 for 14 or 38 weeks. We describe the complete lysosomal degradation of atrophying tubular epithelial cells. During two experiments of 14 and 38 weeks duration, respectively, a total of 15 Beagle dogs received 0.8 g K2HPO4/kg body weight daily with their food. All dogs were examined clinically at regular intervals. Renal biopsies were taken in the fourth week from beagles of the 14-week study. Results were compared with those of control dogs. At the end of the experiments the animals were killed and necropsies done. Different stains and histochemical reactions were applied to paraffin sections of the kidneys. Acid phosphatase and β-glucuronidase were found on cryostat sections. Kidneys fixed by perfusion of five Beagles from the 38-week study and three Beagles of the 14-week study, and from five control dogs, were examined electron microscopically. Ultrahistochemically, acid phosphatase was demonstrated. Clinically, the dogs in both experiments vomited, were cachectic, and had elevated creatinine and blood urea nitrogen. Morphologically, qualitatively identical changes were seen, but the renal damage was most marked at 38 weeks. There were disseminated tubular atrophy (usually of the proximal tubules), focal scar tissue and nephrocalcinosis. The following pathogenesis was established for the lesions of the proximal tubule: Tubular atrophy begins with loss of differentiation of epithelial cells. Enzyme histochemistry, ultrahistochemistry and electron microscopy show an increase in autophagic vacuoles and autophagolysosomes. The lysosomal bodies showing fusion enclose large parts of the cytoplasm as the process continues. Complete lysosomal degradation of epithelial cells and extrusion of large lysosomes into the tubular lumen follow. After complete enzymatic digestion of the intratubular detritus, the residue is empty, convoluted and collapsed tubular basement membrane. Atrophic tubular epithelial cells have many organelle-free zones at their base, which contain fine filamentous material resembling that of the basement membrane. The degradation processes described here may explain why clinically the urinary sediment contains few cylinders and epithelial cells and why proteinuria decreases significantly toward the end of the experiment. So far, it is not clear whether the tubular basement membrane is synthesized by the tubular cells, by fibroblasts or by both cell types. The presence of basement membrane-like material in tubular epithelial cells and in parietal epithelial cells of the glomerulus favors the view that epithelial cells produce the basement membranes and that increased production of basement membrane-like material is a sign of loss of differentiation.

scholarly journals cDNA Cloning of the Basement Membrane Chondroitin Sulfate Proteoglycan Core Protein, Bamacan: A Five Domain Structure Including Coiled-Coil Motifs

1997 ◽  
Vol 136 (2) ◽  
pp. 433-444 ◽  
Author(s):  
Rong-Rong Wu ◽  
John R. Couchman
Keyword(s):  
Basement Membrane ◽  
Chondroitin Sulfate ◽  
Core Protein ◽  
Coiled Coil ◽  
Basement Membranes ◽  
Cdna Clones ◽  
Molecular Architecture ◽  
Unusual Structure ◽  
Extracellular Matrix Molecule

Basement membranes contain several proteoglycans, and those bearing heparan sulfate glycosaminoglycans such as perlecan and agrin usually predominate. Most mammalian basement membranes also contain chondroitin sulfate, and a core protein, bamacan, has been partially characterized. We have now obtained cDNA clones encoding the entire bamacan core protein of Mr = 138 kD, which reveal a five domain, head-rod-tail configuration. The head and tail are potentially globular, while the central large rod probably forms coiled-coil structures, with one large central and several very short interruptions. This molecular architecture is novel for an extracellular matrix molecule, but it resembles that of a group of intracellular proteins, including some proposed to stabilize the mitotic chromosome scaffold. We have previously proposed a similar stabilizing role for bamacan in the basement membrane matrix. The protein sequence has low overall homology, apart from very small NH2- and COOH-terminal motifs. At the junctions between the distal globular domains and the coiled-coil regions lie glycosylation sites, with up to three N-linked oligosaccharides and probably three chondroitin chains. Three other Ser-Gly dipeptides are unfavorable for substitution. Fusion protein antibodies stained basement membranes in a pattern commensurate with bamacan, and they also Western blotted bamacan core protein from rat L2 cell cultures. The antibodies could also specifically immunoprecipitate an in vitro transcription/translation product from a full-length bamacan cDNA. The unusual structure of this proteoglycan is indicative of specific functional roles in basement membrane physiology, commensurate with its distinct expression in development and changes in disease models.

scholarly journals Extracellular matrix of lymphoid tissues in the chick.

1989 ◽  
Vol 37 (5) ◽  
pp. 757-763 ◽  
Author(s):  
A Colombatti ◽  
A Poletti ◽  
A Carbone ◽  
D Volpin ◽  
G M Bressan
Keyword(s):  
Extracellular Matrix ◽  
Epithelial Cells ◽  
Basement Membrane ◽  
Basement Membranes ◽  
Intestinal Lumen ◽  
Lymphoid Tissues ◽  
White Pulp ◽  
Extracellular Matrix Components ◽  
Coarse Fibers ◽  
Distribution Of Components

We describe the immunohistochemical distribution of components of the extracellular matrix of the chick lymphoid system. In the thymus, basement membranes of epithelial cells bordering the lobules were intensely stained by laminin antibodies; fibronectin antibodies labeled the capsule and the septal matrix, and similar reactivity was seen with tropoelastin and gp 115 antibodies. No positivity was detected with any of the antibodies within the cortical parenchymal cells. Laminin was not detected in the medullary parenchyma, whereas fibronectin was present as coarse fibers. Tropoelastin and gp 115 appeared as a finer and more diffuse meshwork. In the bursa, laminin antibodies outlined the epithelial cells separating the cortex from the medulla. Fibronectin, tropoelastin, and gp 115 antibody stained the interfollicular septa and the cortical matrix, although to a different extent. Laminin was also detected in association with the interfollicular epithelium (IFE) basement membrane, whereas no staining was found underneath the follicle-associated epithelium (FAE). FAE cells not only lack a proper basement membrane but are also not separated from medullary lymphocytes by any of the other extracellular matrix components were investigated. Consequently, medullary lymphocytes are not sequestered, and can come easily into contact with antigens present in the intestinal lumen. All four antibodies stained the spleen capsule and spleen blood vessels, tropoelastin and gp 115 antibodies giving the strongest reactivity. A fine trabecular staining pattern was detected with gp 115 antibodies in the white pulp.

123 GENISTEIN TRANSPORT ACROSS THE BOVINE OVIDUCT EPITHELIUM

2013 ◽  
Vol 25 (1) ◽  
pp. 208 ◽  
Author(s):  
C. Simintiras ◽  
F. L. Courts ◽  
R. G. Sturmey
Keyword(s):  
Epithelial Cells ◽  
Room Temperature ◽  
Cell Monolayer ◽  
Barrier Properties ◽  
Apparent Permeability ◽  
Mammalian Embryo ◽  
Significant Difference ◽  
Apical Compartment ◽  
Oviduct Epithelium

The oviduct plays a vital role in regulating the environment surrounding the gametes and early mammalian embryo. However, the permeability of the oviduct to circulating dietary-derived compounds remains relatively unknown. The present study has investigated the barrier properties of the oviduct epithelium in vitro to the movement of genistein, a soya isoflavone and analogue of 17β-estradiol that naturally occurs in the diet and has been reported to exert teratogenic effects. Bovine oviduct epithelial cells (BOECs) were isolated from abattoir-derived reproductive tracts at the mid-luetal phase. The purity of the cell isolate was confirmed using flow cytometry to determine the proportion of cells that expressed CK18 (epithelia), and Vimentin (fibroblasts). Cells were seeded at a density of 1 × 106 mL–1 on polyethylene terephthalate transwell porous supports (Corning) and maintained between two media-filled chambers for 10 to 12 days, until they formed a polarised confluent monolayer, as confirmed by transepithelial resistance (TEER) greater than 700 Ωcm2. To assess the rate of transport, genistein was added to the basal compartment at physiologically relevant levels (100 µM) and the apical compartment was sampled at regular time points for 120 mins. The concentration of genistein in the apical and basal media was measured by HPLC. Furthermore, we compared the rate of genistein transport at physiological (39°C) and room temperature to indicate whether transport was temperature dependent. Rates of transport are expressed as mean apparent permeability coefficient and were compared between groups by a Student’s t-test. Genistein crossed the bovine oviduct epithelium at a linear rate that was higher than spontaneous diffusion across a blank membrane support (12.7 × 10–3 cm–2 µM–1 v. 7.32 × 10–3 cm–2 µM–1, n = 4; P = 0.0075). The rate of genistein transport by epithelial cells was unchanged when cells were assayed at room temperature (12.7 × 10–3 cm–2 µM–1 v. 13.11 × 10–3 cm–2 µM–1, n = 3; P = 0.76), respectively. No significant difference in the directionality of transport was found. Furthermore, TEER was maintained at approximately 700 Ωcm2, indicating that the cells remained confluent for the duration of the experiment. These data suggest that the bovine oviduct epithelial cell monolayer facilitates genistein movement from the basal to the apical compartment in vitro. Furthermore, the observation that the rate of transport is unchanged by temperature suggests a passive, trans-cellular, physicochemical mechanism, rather than an active biological process. Regardless of the mechanism, the oviduct epithelium is permeable to genistein, and may even facilitate its transport into the lumen, suggesting that gametes and early embryos could be exposed to this compound in vivo. This is relevant given the previously reported finding indicating that this naturally occurring dietary isoflavone has detrimental effects on early development (Newbold et al. 2001 Cancer Res. 61, 4325–4328). Furthermore, the results demonstrate the potential use of this epithelial model in characterising the transport or barrier properties of the oviduct epithelium towards a range of circulating xenobiotics.

scholarly journals Human Bronchial Epithelial Cells Secrete Laminin 5, Express Hemidesmosomal Proteins, and Assemble Hemidesmosomes

2000 ◽  
Vol 48 (4) ◽  
pp. 535-544 ◽  
Author(s):  
Peter H. Michelson ◽  
Margaret Tigue ◽  
Jonathan C.R. Jones
Keyword(s):  
Epithelial Cell ◽  
Epithelial Cells ◽  
Basement Membrane ◽  
Cell Monolayer ◽  
Bronchial Epithelial Cells ◽  
Basal Cells ◽  
Bronchial Epithelial ◽  
Laminin 5 ◽  
Α6β4 Integrin

Epithelial cells attach to the basement membrane through adhesive contacts between the basal cells of the epithelium and the proteins of the extracellular matrix (ECM). The hemidesmosome (HD) is a specialized cell-ECM contact, that mediates the attachment of the epithelial cell basal surface to the ECM. In bronchial epithelial cells, the protein components that constitute the HD have not been demonstrated. Using immunohistochemical techniques, we determined that normal human bronchial epithelial (NHBE) cells express the HD cell surface integrin α6β4 and produce laminin 5, the ECM protein associated with HDs. Furthermore, expression of the HD-associated structural proteins, bullous pemphigoid antigens 1 (BPAG 1) and 2 (BPAG 2), was demonstrated in NHBE cells by immunofluorescence microscopy and immunoblot analyses. In addition, we confirmed the presence of laminin 5 in the basement membrane (BM) of bronchial epithelial biopsy specimens and of BP230, BP180, and the α6β4 integrin heterodimer at the site of bronchial epithelial cell-ECM interaction in vivo. Finally, using electron microscopy, we were able to demonstrate intact HDs in a glutaraldehyde-fixed NHBE cell monolayer. These findings suggest that bronchial epithelium forms HDs and that the laminin 5-α6β4 integrin interaction may be important in stabilizing epithelial cell adhesion to the BM in the lung.

scholarly journals Electrical currents flow out of domes formed by cultured epithelial cells.

1984 ◽  
Vol 99 (4) ◽  
pp. 1541-1544 ◽  
Author(s):  
K Sugahara ◽  
J H Caldwell ◽  
R J Mason
Keyword(s):  
Epithelial Cell ◽  
Epithelial Cells ◽  
Active Transport ◽  
Cell Monolayer ◽  
Extracellular Fluid ◽  
Outward Current ◽  
Alveolar Epithelial Cells ◽  
Transepithelial Transport ◽  
Alveolar Type

Domes are localized areas of fluid accumulation between a cultured epithelial cell monolayer and the impermeable substratum on which the cells are cultured in vitro. Dome formation has been documented in a variety of epithelial cell lines that retain their transepithelial transport properties in vitro. However, it is not known whether domes are predominantly areas of specific active transport, or, alternatively, are predominantly areas of relative weak attachment to the culture surface. In the present study we adapted a vibrating microelectrode, which can detect small currents flowing in extracellular fluid, to determine if current was flowing into or out of domes and thereby to determine if domes were specialized areas of active transport. We used alveolar type II cells as the main epithelial cell type because they readily form domes in vitro and because they transport sodium from the apical to the basal surface. We found that electrical current flowed out of domes. The direction of the current was independent of the size of a dome, of the age of an individual dome, and of the number of days in primary culture for alveolar epithelial cells. This current was inhibited by amiloride and ouabain and was dependent on sodium in the medium. We made similar observations (outward current from domes which is blocked by amiloride and by sodium substitution) with domes formed by the Madin-Darby canine kidney cell line. The data support the hypothesis that sodium is transported across the entire monolayer and leaks back mainly through the domes. We conclude that domes in epithelial monolayers are not predominantly special sites of active transport but are more likely simply areas of weak attachment to the substratum.

Laminin α-chain expression and basement membrane formation by MLE-15 respiratory epithelial cells

2002 ◽  
Vol 282 (5) ◽  
pp. L1004-L1011 ◽  
Author(s):  
Nguyet M. Nguyen ◽  
Yushi Bai ◽  
Katsumi Mochitate ◽  
Robert M. Senior
Keyword(s):  
Epithelial Cells ◽  
Basement Membrane ◽  
Critical Role ◽  
Basement Membranes ◽  
Alveolar Type ◽  
Type Ii Cells ◽  
Type Ii ◽  
Respiratory Epithelial Cells ◽  
Alveolar Type Ii Cells ◽  
Respiratory Epithelial

Basement membranes have a critical role in alveolar structure and function. Alveolar type II cells make basement membrane constituents, including laminin, but relatively little is known about the production of basement membrane proteins by murine alveolar type II cells and a convenient system is not available to study basement membrane production by murine alveolar type II cells. To facilitate study of basement membrane production, with particular focus on laminin chains, we examined transformed murine distal respiratory epithelial cells (MLE-15), which have many structural and biochemical features of alveolar type II cells. We found that MLE-15 cells produce laminin-α5, a trace amount of laminin-α3, laminins-β1 and -γ1, type IV collagen, and perlecan. Transforming growth factor-β1 significantly induces expression of laminin-α1. When grown on a fibroblast-embedded collagen gel, MLE-15 cells assemble a basement membrane-like layer containing laminin-α5. These findings indicate that MLE-15 cells will be useful in modeling basement membrane production and assembly by alveolar type II cells.

Studies in vitro of effects of steroid hormones and the blastocyst on endometrial function in the sheep

1992 ◽  
Vol 4 (3) ◽  
pp. 275 ◽  
Author(s):  
LA Salamonsen ◽  
RA Cherny ◽  
JK Findlay
Keyword(s):  
Epithelial Cells ◽  
Steroid Hormones ◽  
Cell Types ◽  
Basement Membranes ◽  
Two Dimensional Gel Electrophoresis ◽  
In Vitro Techniques ◽  
Endometrial Cells ◽  
Matrix Metalloproteinase 3 ◽  
Prostaglandin Synthase

Normal endometrial function is a result of regulation by the combination of ovarian steroids and local agents arising from within the embryo-maternal unit. We have used in vitro techniques to examine the role of steroid hormones and ovine trophoblast interferon on endometrial function in the ewe. Immunolocalization of oestrogen receptors in endometrial tissue demonstrated marked changes throughout the cycle and in early pregnancy with maximal concentrations during the follicular and very early luteal phases. Protein secretion from highly purified cultured ovine stromal and epithelial endometrial cells, and the direction of secretion from polarized epithelial cells, has been examined by incorporation of [35S]methionine and by one- and two-dimensional gel electrophoresis. Protein synthesis is greater in stromal than in epithelial cells and more protein is secreted apically than basally from epithelial cells. A number of common and some different proteins are secreted by the two cell types. One secreted protein is matrix metalloproteinase-3 (stromelysin) which degrades components of basement membranes. Ovine trophoblast interferon attenuates the production of prostaglandins from ovine endometrial cells but its action is not by an effect on localization or concentration of the enzyme prostaglandin synthase or on expression of the gene for prostaglandin synthase. Such studies in vitro contribute to our understanding of how the endometrium is prepared for implantation.

scholarly journals Expression of extracellular matrix components is regulated by substratum.

1990 ◽  
Vol 110 (4) ◽  
pp. 1405-1415 ◽  
Author(s):  
C H Streuli ◽  
M J Bissell
Keyword(s):  
Extracellular Matrix ◽  
Epithelial Cells ◽  
Basement Membrane ◽  
Type I Collagen ◽  
Basement Membranes ◽  
Type I ◽  
Collagen Gels ◽  
Extracellular Matrix Components ◽  
Matrix Components ◽  
Cells Cultured

Reconstituted basement membranes and extracellular matrices have been demonstrated to affect, positively and dramatically, the production of milk proteins in cultured mammary epithelial cells. Here we show that both the expression and the deposition of extracellular matrix components themselves are regulated by substratum. The steady-state levels of the laminin, type IV collagen, and fibronectin mRNAs in mammary epithelial cells cultured on plastic dishes and on type I collagen gels have been examined, as has the ability of these cells to synthesize, secrete, and deposit laminin and other, extracellular matrix proteins. We demonstrate de novo synthesis of a basement membrane by cells cultured on type I collagen gels which have been floated into the medium. Expression of the mRNA and proteins of basement membranes, however, are quite low in these cultures. In contrast, the levels of laminin, type IV collagen, and fibronectin mRNAs are highest in cells cultured on plastic surfaces, where no basement membrane is deposited. It is suggested that the interaction between epithelial cells and both basement membrane and stromally derived matrices exerts a negative influence on the expression of mRNA for extracellular matrix components. In addition, we show that the capacity for lactational differentiation correlates with conditions that favor the deposition of a continuous basement membrane, and argue that the interaction between specialized epithelial cells and stroma enables them to create their own microenvironment for accurate signal transduction and phenotypic function.

Sign in / Sign up

Export Citation Format

Share Document