Lectin Staining of Peritoneal Mesothelial Cells in Vitro

A survey of lectin-binding specificities present on rodent and human mesothelial cells propagated and maintained in tissue culture was made using fluorescein isothiocynate conjugated (FITC) lectins. Rodent and human cells exhibited cell-associated fluorescence following exposure to the FITC-Iectins from C. ensiformis, T. vulgaris, A. hypogaea, E. cristagalli and B. simplicifolia, but not with lectins from G. max and D. biflorus. Rodent cells were also positive for FITC-M. pomifera lectin binding. Human, but not rodent, cells were positive for FITC T. purpureas lectin binding. Exposure of rabbit mesothelial cells in vitro to FITC-Iectins that bound to the cell surface resulted in the appearance of discrete loci of putatively intracellular fluorescence. Exposure of cells to ferritin-Iabelled T. vulgaris lectin at 37°C for as little as 7.5 minutes resulted in the appearance of ferritin-size particles in intracellular vesicles. These results demonstrate 1. the presence of lectinbinding sites in and on peritoneal mesothelial cells from rodents and humans and 2. a possible role of such sites in mediating the entry of lectin-Iike endogenous molecules into the vacuolar apparatus of these cells.

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The Mesothelial Cell as a Non-Thrombogenic Surface

Thrombosis and Haemostasis ◽

10.1055/s-0038-1661149 ◽

1984 ◽

Vol 52 (02) ◽

pp. 102-104 ◽

Cited By ~ 25

Author(s):

L J Nicholson ◽

J M F Clarke ◽

R M Pittilo ◽

S J Machin ◽

N Woolf

Keyword(s):

Electron Microscopy ◽

Scanning Electron Microscopy ◽

Blood Flow ◽

Cell Surface ◽

Mesothelial Cell ◽

Mesothelial Cells ◽

Plastic Film ◽

In Vitro System ◽

Scanning Electron

SummaryA technique for harvesting mesothelial cells is described. This entails collagenase digestion of omentum after which the cells can be cultured. The technique has been developed using the rat, but has also been successfully applied to human tissue. Cultured rat mesothelial cells obtained in this way have been examined by scanning electron microscopy. Rat mesothelial cells grown on plastic film have been exposed to blood in an in vitro system using a Baumgartner chamber and have been demonstrated to support blood flow. No adhering platelets were observed on the mesothelial cell surface. Fibroblasts similarily exposed to blood as a control were washed off the plastic.

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Neural induction and the structure of the target cell surface

Development ◽

10.1242/dev.70.1.171 ◽

1982 ◽

Vol 70 (1) ◽

pp. 171-187

Author(s):

A. M. Duprat ◽

L. Gualandris ◽

P. Rouge

Keyword(s):

Cell Surface ◽

Structural Integrity ◽

Neural Induction ◽

Active Role ◽

Target Cells ◽

Similar Treatment ◽

Structural Modifications ◽

Pleurodeles Waltl

Lectins (SBA and PSA) were used to provoke crowding and structural modifications of the presumptive ectoderm cell surface in order to investigate the role of the membrane organization of the competent target cells in neural induction. Are specific characteristics of the cell surface essential for this phenomenon to occur? From amphibian gastrulae, it is possible to obtain neural induction in vitro by association of presumptive ectoderm (target cells) with chordamesoderm (inductor tissue): 4 h of contact is sufficient in Pleurodeles waltl for transmission of the inductive signal. Very quickly, the treatment of the normal ectoderm by lectins (SBA-FITC or PSA-FITC) provoked surface modifications. Lectin-treatment (50 µg ml1−, 30 min) of presumptive ectoderm did not result in any neural induction. Lectin-treatment (50 µg ml1−, 30 min) of presumptive ectoderm previous to its association with the natural inductor for 4 h, disturbed the phenomenon: no induction. Similar treatment followed by association with the inductor for 24 h: induction. Treatment of SBA or PSA with their respective hapten inhibitors prior to addition to ectodermal cells completely blocked the suppressive effects on induction. The structural integrity of the membrane of competent target cells is necessary for neural induction to occur. The cell membrane could thus play, directly or indirectly, an active role in the specificity of this process

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Mesothelial Cells

Peritoneal Dialysis International ◽

10.1177/089686080702702s19 ◽

2007 ◽

Vol 27 (2_suppl) ◽

pp. 110-115 ◽

Cited By ~ 5

Author(s):

Susan Yung ◽

Chan Tak Mao

Keyword(s):

Mesothelial Cell ◽

In Vitro Models ◽

Mesothelial Cells ◽

Dynamic Membrane ◽

Peritoneal Mesothelial Cells ◽

Immunohistochemical Markers ◽

Vitro Model ◽

And Function

♦ Background The introduction of peritoneal dialysis (PD) as a modality of renal replacement therapy has provoked much interest in the biology of the peritoneal mesothelial cell. Mesothelial cells isolated from omental tissue have immunohistochemical markers that are identical to those of mesothelial stem cells, and omental mesothelial cells can be cultivated in vitro to study changes to their biologic functions in the setting of PD. ♦ Method The present article describes the structure and function of mesothelial cells in the normal peritoneum and details the morphologic changes that occur after the introduction of PD. Furthermore, this article reviews the literature of mesothelial cell culture and the limitations of in vitro studies. ♦ Results The mesothelium is now considered to be a dynamic membrane that plays a pivotal role in the homeostasis of the peritoneal cavity, contributing to the control of fluid and solute transport, inflammation, and wound healing. These functional properties of the mesothelium are compromised in the setting of PD. Cultures of peritoneal mesothelial cells from omental tissue provide a relevant in vitro model that allows researchers to assess specific molecular pathways of disease in a distinct population of cells. Structural and functional attributes of mesothelial cells are discussed in relation to long-term culture, proliferation potential, age of tissue donor, use of human or animal in vitro models, and how the foregoing factors may influence in vitro data. ♦ Conclusions The ability to propagate mesothelial cells in culture has resulted, over the past two decades, in an explosion of mesothelial cell research pertaining to PD and peritoneal disorders. Independent researchers have highlighted the potential use of mesothelial cells as targets for gene therapy or transplantation in the search to provide therapeutic strategies for the preservation of the mesothelium during chemical or bacterial injury.

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MiR-26a-5p inhibits GSK3β expression and promotes cardiac hypertrophy in vitro

PeerJ ◽

10.7717/peerj.10371 ◽

2020 ◽

Vol 8 ◽

pp. e10371

Author(s):

Liqun Tang ◽

Jianhong Xie ◽

Xiaoqin Yu ◽

Yangyang Zheng

Keyword(s):

Cell Surface ◽

Cardiac Hypertrophy ◽

Surface Area ◽

Myocardial Cell ◽

Immunofluorescence Staining ◽

Cell Surface Area ◽

Direct Target

Background The role of miR-26a-5p expression in cardiac hypertrophy remains unclear. Herein, the effect of miR-26a-5p on cardiac hypertrophy was investigated using phenylephrine (PE)-induced cardiac hypertrophy in vitro and in a rat model of hypertension-induced hypertrophy in vivo. Methods The PE-induced cardiac hypertrophy models in vitro and vivo were established. To investigate the effect of miR-26a-5p activation on autophagy, the protein expression of autophagosome marker (LC3) and p62 was detected by western blot analysis. To explore the effect of miR-26a-5p activation on cardiac hypertrophy, the relative mRNA expression of cardiac hypertrophy related mark GSK3β was detected by qRT-PCR in vitro and vivo. In addition, immunofluorescence staining was used to detect cardiac hypertrophy related mark α-actinin. The cell surface area was measured by immunofluorescence staining. The direct target relationship between miR-26a-5p and GSK3β was confirmed by dual luciferase report. Results MiR-26a-5p was highly expressed in PE-induced cardiac hypertrophy. MiR-26a-5p promoted LC3II and decreased p62 expression in PE-induced cardiac hypertrophy in the presence or absence of lysosomal inhibitor. Furthermore, miR-26a-5p significantly inhibited GSK3β expression in vitro and in vivo. Dual luciferase report results confirmed that miR-26a-5p could directly target GSK3β. GSK3β overexpression significantly reversed the expression of cardiac hypertrophy-related markers including ANP, ACTA1 and MYH7. Immunofluorescence staining results demonstrated that miR-26a-5p promoted cardiac hypertrophy related protein α-actinin expression, and increased cell surface area in vitro and in vivo. Conclusion Our study revealed that miR-26a-5p promotes myocardial cell autophagy activation and cardiac hypertrophy by regulating GSK3β, which needs further research.

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Role of peritoneal mesothelial cells in peritonitis

British Journal of Surgery ◽

10.1002/bjs.4373 ◽

2003 ◽

Vol 90 (10) ◽

pp. 1187-1194 ◽

Cited By ~ 36

Author(s):

V. Yao ◽

C. Platell ◽

J. C. Hall

Keyword(s):

Mesothelial Cells ◽

Peritoneal Mesothelial Cells

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Role of oxygen derivatives in the cytotoxicity and DNA damage produced by asbestos on rat pleural mesothelial cells in vitro

Carcinogenesis ◽

10.1093/carcin/15.6.1251 ◽

1994 ◽

Vol 15 (6) ◽

pp. 1251-1255 ◽

Cited By ~ 33

Author(s):

Hang Ying Dong ◽

Annie Buard ◽

Annie Renier ◽

Francoise Lévy ◽

Laure Saint-Etienne ◽

...

Keyword(s):

Dna Damage ◽

Mesothelial Cells ◽

Pleural Mesothelial Cells

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The Role of Tgf-β in Growth Inhibition of Peritoneal Mesothelial Cells in High-Glucose Dialysate

Peritoneal Dialysis International ◽

10.1177/089686089501507s13 ◽

1995 ◽

Vol 15 (7_suppl) ◽

pp. 93-95 ◽

Cited By ~ 19

Author(s):

Kazuo Kumano ◽

Masuhiro Shimoda ◽

Toru Hyodo ◽

Tadasu Sakai

Keyword(s):

Growth Inhibition ◽

High Glucose ◽

Mesothelial Cells ◽

Peritoneal Mesothelial Cells

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Compartmentalization of protein traffic in insulin-sensitive cells

AJP Endocrinology and Metabolism ◽

10.1152/ajpendo.1996.271.1.e1 ◽

1996 ◽

Vol 271 (1) ◽

pp. E1-E14 ◽

Cited By ~ 16

Author(s):

K. V. Kandror ◽

P. F. Pilch

Keyword(s):

Cell Surface ◽

Physiological Role ◽

Fat Cells ◽

Protein Traffic ◽

Glut 1 ◽

Glut 4 ◽

Factor Ii ◽

Nutritional Needs

Insulin-sensitive cells, adipocytes and myocytes, translocate a number of intracellular proteins to the cell surface in response to insulin. Among these proteins are glucose transporters 1 and 4 (GLUT-1 and GLUT-4, respectively), receptors for insulin-like growth factor II (IGF-II)/mannose 6-phosphate (Man-6-P) and transferrin, the aminopeptidase gp 160, caveolin, and a few others. In the case of insulin-activated glucose transport, this translocation has been proven to be the major, if not the only regulatory mechanism of this process. It seems likely that the cell surface recruitment of the IGF-II/Man-6-P and transferrin receptors also serves the nutritional needs of cells, whereas the physiological role of the aminopeptidase gp160 remains uncertain. Analysis of the compartmentalization and trafficking pathways of translocatable proteins in fat cells identified more than one population of recycling vesicles, although all have identical sedimentation coefficients and buoyant densities in vitro. GLUT-4-containing vesicles include essentially all the intracellular GLUT-4, gp160, and the acutely recycling populations of receptors for IGF-II/Man-6-P and transferrin. Besides these proteins, which can be considered as vesicle “cargo”, GLUT-4-containing vesicles have other components, like secretory carrier-associated membrane proteins (SCAMP), Rab(s), and vesicle-associated membrane protein (VAMP)/cellubrevin, which are ubiquitous to secretory vesicles and granules from different tissues. GLUT-1 and caveolin are excluded from GLUT-4-containing vesicles and form different vesicular populations of unknown polypeptide composition. In skeletal muscle, two independent populations of GLUT-4-containing vesicles are found, insulin sensitive and exercise sensitive, which explains the additive effect of insulin and exercise on glucose uptake. Both vesicular populations are similar to each other and to analogous vesicles in fat cells.

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In Vivo and In Vitro Characterization of Mesothelial Lipid Inclusions

Peritoneal Dialysis International ◽

10.1177/089686089101100304 ◽

1991 ◽

Vol 11 (3) ◽

pp. 207-212 ◽

Cited By ~ 7

Author(s):

J. Thomas Hjelle ◽

Barbara T. Golinska ◽

Diane C. Waters ◽

Kevin R. Steidley ◽

David R. McCarroll ◽

...

Keyword(s):

Electron Microscopy ◽

Culture Media ◽

Morphological Characteristics ◽

Mesothelial Cells ◽

Lipid Bodies ◽

Lamellar Bodies ◽

Peritoneal Mesothelial Cells ◽

Lipid Inclusions

The nature of intracytoplasmic lipid inclusions found in cultured rabbit and rat peritoneal mesothelial cells was examined by ultrastructural and biochemical techniques. Transmission electron microscopy also demonstrated extracellular release of these lipid bodies. Differential fixation with tannic acid revealed 2 types of inclusions, lamellated (lamellar bodies) and nonlamellated (homogeneous). The lamellar bodies were found near or in the Golgi apparatus and on the cell surface where occasionally they were observed in exocytotic pouches. The homogeneous inclusions were the predominant species being found primarily intracellularly. Lipid bodies obtained from the culture media over the cells displayed on electron microscopy the same morphological characteristics as those seen intracellularly. Exposure of confluent cultures of mesothelial cells to the vital lipid stain Nile Red caused the appearance of intensely fluorescent droplets in or on the cells at wave lengths consistent with staining for phosphatidylcholine-rich vesicles. Incubation of the cells with r4C)-choline an d subsequent analysis of phospholipid formation revealed high rates of r4C)-phosphatidylcholine addition to both intra and extracellular lipid pools. Taken together, mesothelial cells exhibit lipid bodies similar in ultrastructure to the surfactant containing organelles of Type II pneumocytes.

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