Glycerol Toxicity for Human Peritoneal Mesothelial Cells in Culture: Comparison with Glucose

1994 ◽  
Vol 17 (5) ◽  
pp. 252-260 ◽  
Author(s):  
J. Witowski ◽  
J. Knapowski
Keyword(s):  
Mesothelial Cell ◽  
Cellular Protein ◽  
Mesothelial Cells ◽  
Dependent Manner ◽  
Peritoneal Mesothelial Cells ◽  
Proline Incorporation ◽  
Human Peritoneal Mesothelial Cells ◽  
Dose Dependent Decrease ◽  
Dose Dependent

Glycerol has been proposed as a substitute osmotic agent for glucose in peritoneal dialysis fluids. We have compared the effect of glycerol and glucose on the function of human peritoneal mesothelial cells (HPMC) in vitro. The viability of HPMC was not affected by glycerol (up to 250 mM), whereas it was reduced by glucose in a time- and dose-dependent manner, as assessed by the LDH release. Although the incubation of HPMC with glycerol induced a dose-dependent decrease in HPMC proliferation, the effect was significantly less inhibitory than that produced by glucose. In HPMC treated with 90 mM of glycerol or glucose the incorporation of [3H]-thymidine had reached 79.0±19.3% and 55.3+4.0% of the control (p<0.05 and p<0.01), respectively. As measured by the [methyl-14C]-choline incorporation, the intracellular amount of newly synthesized phospholipids was reduced from (cpm/μg cellular protein) 147±58 in control HPMC to 59+15 in cells exposed to 90 mM of glucose (p<0.01), but not affected by glycerol (163±65). On the other hand, both glycerol and glucose (90 mM) decreased the synthesis of proteins (as assessed by the [3H]-proline incorporation) and interfered with potassium (86Rb) transport mechanisms in HPMC. Our data suggest that there exist some possibly advantageous aspects of glycerol as far as mesothelial cell biocompatibility profile is concerned.

Mesothelial Cell Adherence to Vascular Prostheses and Their Subsequent Growth In Vitro

1994 ◽  
Vol 3 (1) ◽  
pp. 41-48 ◽  
Author(s):  
Apollo Pronk ◽  
Arthur A.G.M. Hoynck Van Papendrecht ◽  
Piet Leguit ◽  
Henri A. Verbrugh ◽  
Roel P.A.J. Verkooyen ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Cell Attachment ◽  
Mesothelial Cell ◽  
Mesothelial Cells ◽  
Subsequent Growth ◽  
Vascular Prostheses ◽  
Peritoneal Mesothelial Cells ◽  
Viable Cells ◽  
Human Peritoneal Mesothelial Cells

Cell seeding may decrease the thrombogenicity of implanted vascular grafts, but its application is hampered by the limited availability of autologous endothelial cells. Human peritoneal mesothelial cells have blood flow supporting qualities and are readily available. This study investigated the adherence of mesothelial cells to vascular prostheses and their subsequent growth in vitro. Circular pieces of various vascular prosthetic materials were seeded with 51Chromium-labeled mesothelial and endothelial cells and left for either 5, 15, 30, 60, and 120 minutes. The unattached cells were removed and the degree of cell attachment was measured. The number of mesothelial cells to Dacron increased during the first 60 min up to 35.2 % of the seeded inoculum whereafter a plateau was reached. Scanning electron microscopy showed spreaded mesothelial cells adherent to the Dacron fibers. A significant increase in adherence was observed after preincubation of Dacron with 10 μg/mL fibronectin, but no improvement was found after preincubation with human serum albumin or gelatin. Mesothelial cells adhered better to Gelcoated than to Gelsealed or plain Dacron. The adherence of mesothelial cells to ePTFE (Teflon) was significantly poorer. No significant differences in adherence were found between mesothelial and endothelial cells. Mesothelial cell growth on Dacron resulted in a modest increase in the number of viable cells during 27 days, which implies biocompatibility of Dacron and mesothelial cells in vitro.

scholarly journals miRNA589 Regulates Epithelial-Mesenchymal Transition in Human Peritoneal Mesothelial Cells

2012 ◽  
Vol 2012 ◽  
pp. 1-6 ◽  
Author(s):  
Ke Zhang ◽  
Hao Zhang ◽  
Xun Zhou ◽  
Wen-bin Tang ◽  
Li Xiao ◽  
...  
Keyword(s):  
Epithelial Mesenchymal Transition ◽  
Mesothelial Cell ◽  
Mesothelial Cells ◽  
Control Group ◽  
Peritoneal Fibrosis ◽  
Mesenchymal Transition ◽  
Peritoneal Mesothelial Cells ◽  
Human Peritoneal Mesothelial Cells ◽  
E Cadherin

Background. microRNA (miRNA, miR) are thought to interact with multiple mRNAs which are involved in the EMT process. But the role of miRNAs in peritoneal fibrosis has remained unknown.Objective. To determine if miRNA589 regulates the EMT induced by TGFβ1 in human peritoneal mesothelial cell line (HMrSV5 cells).Methods. 1. Level of miR589 was detected in both human peritoneal mesothelial cells (HPMCs) isolated from continuous ambulatory peritoneal dialysis (CAPD) patients’ effluent and HMrSV5 cells treated with or without TGFβ1. 2. HMrSV5 cells were divided into three groups: control group, TGFβ1 group, and pre-miR-589+TGFβ1 group. The level of miRNA589 was determined by realtime PCR. The expressions of ZO-1, vimentin, and E-cadherin in HPMCs were detected, respectively.Results. Decreased level of miRNA589 was obtained in either HPMCs of long-term CAPD patients or HMrSV5 cells treated with TGFβ1. In vitro, TGFβ1 led to upregulation of vimentin and downregulation of ZO-1 as well as E-cadherin in HMrSV5 cells, which suggested EMT, was induced. The changes were accompanied with notably decreased level of miRNA589 in HMrSV5 cells treated with TGFβ1. Overexpression of miRNA589 by transfection with pre-miRNA589 partially reversed these EMT changes.Conclusion. miRNA589 mediates TGFβ1 induced EMT in human peritoneal mesothelial cells.

Interleukin-1β Stimulates Glucose Uptake of Human Peritoneal Mesothelial Cells In Vitro

1996 ◽  
Vol 16 (1_suppl) ◽  
pp. 58-60 ◽  
Author(s):  
Michael Kruse ◽  
Arezki Mahiout ◽  
Volker Kliem ◽  
Peter Kurz ◽  
Karl-Martin Koch ◽  
...  
Keyword(s):  
Glucose Uptake ◽  
Glucose Transporter ◽  
Cytochalasin B ◽  
Interleukin 1 ◽  
Glucose Transporters ◽  
Mesothelial Cells ◽  
Dependent Manner ◽  
Peritoneal Mesothelial Cells ◽  
Human Peritoneal Mesothelial Cells

To investigate whether the glucose uptake (GU) of human peritoneal mesothelial cells (HPMC) is mediated by glucose transporters and whether this uptake is influenced by interleukin 1–β (IL-1β), we measured 2-deoxy-(3H)-GU of HPMC in vitro, after exposing the cells for different times (two and 12 hours) to increasing concentrations (0.1, 1.0, and 2.0 ng/mL) of IL-1 β. To exclude a noncarrier-mediated transport, GU was also tested in the presence of cytochalasin B. All experiments were performed in triplicate in the cells of two donors. Cytochalasin B inhibits GU of HPMC almost completely. GU of HPMC is not stimulated by insulin. GU is stimulated by IL-1 β in a dose-dependent manner. These data indicate a GU of HPMC, which is mediated by a glucose transporter and stimulated by IL-1 β. The increased uptake of glucose from the dialysate In patients with peritonitis may be mediated by a (cytokineinduced) increased activity of HPMC glucose transporters.

scholarly journals MiR-29b may suppresses peritoneal metastases through inhibition of the mesothelial–mesenchymal transition (MMT) of human peritoneal mesothelial cells

2022 ◽  
Vol 12 (1) ◽  
Author(s):  
Yuki Kimura ◽  
Hideyuki Ohzawa ◽  
Hideyo Miyato ◽  
Yuki Kaneko ◽  
Akira Saito ◽  
...  
Keyword(s):  
Morphological Changes ◽  
Negative Control ◽  
Mesothelial Cells ◽  
Peritoneal Metastases ◽  
Dependent Manner ◽  
Mesenchymal Transition ◽  
Peritoneal Mesothelial Cells ◽  
Human Peritoneal Mesothelial Cells ◽  
Tgf Β1

AbstractPeritoneal dissemination is a major metastatic pathway for gastrointestinal and ovarian malignancies. The miR-29b family is downregulated in peritoneal fluids in patients with peritoneal metastases (PM). We examined the effect of miR-29b on mesothelial cells (MC) which play critical a role in the development of PM through mesothelial-mesenchymal transition (MMT). Human peritoneal mesothelial cells (HPMCs) were isolated from surgically resected omental tissue and MMT induced by stimulation with 10 ng/ml TGF-β1. MiR-29b mimics and negative control miR were transfected by lipofection using RNAiMAX and the effects on the MMT evaluated in vitro. HPMC produced substantial amounts of miR-29b which was markedly inhibited by TGF-β1. TGF-β1 stimulation of HPMC induced morphological changes with decreased expression of E-cadherin and calretinin, and increased expression of vimentin and fibronectin. TGF-β1 also enhanced proliferation and migration of HPMC as well as adhesion of tumor cells in a fibronectin dependent manner. However, all events were strongly abrogated by simultaneous transfection of miR-29b. MiR-29b inhibits TGF-β1 induced MMT and replacement of miR-29b in the peritoneal cavity might be effective to prevent development of PM partly through the effects on MC.

Activation of proteinase-activated receptor-2 in mesothelial cells induces pleural inflammation

2005 ◽  
Vol 288 (4) ◽  
pp. L734-L740 ◽  
Author(s):  
Y.C. Gary Lee ◽  
Darryl A. Knight ◽  
Kirk B. Lane ◽  
Dong Sheng Cheng ◽  
M. Audrey Koay ◽  
...  
Keyword(s):  
Pleural Fluid ◽  
Mesothelial Cell ◽  
Coagulation Factor ◽  
Fold Increase ◽  
Mesothelial Cells ◽  
Dependent Manner ◽  
Pleural Diseases ◽  
Dose Dependent

Pleural inflammation underlies many pleural diseases, but its pathogenesis remains unclear. Proteinase-activated receptor-2 (PAR2) is a novel seven-transmembrane receptor with immunoregulatory roles. We hypothesized that PAR2 is present on mesothelial cells and can induce pleural inflammation. PAR2 was detected by immunohistochemistry in all (19 parietal and 11 visceral) human pleural biopsies examined. In cultured murine mesothelial cells, a specific PAR2-activating peptide (SLIGRL-NH2) at 10, 100, and 1,000 μM stimulated a 3-, 42-, and 1,330-fold increase of macrophage inflammatory protein (MIP)-2 release relative to medium control, respectively ( P < 0.05 all) and a 2-, 32-, and 75-fold rise over the control peptide (LSIGRL-NH2, P < 0.05 all). A similar pattern was seen for TNF-α release. Known physiological activators of PAR2, tryptase, trypsin, and coagulation factor Xa, also stimulated dose-dependent MIP-2 release from mesothelial cells in vitro. Dexamethasone inhibited the PAR2-mediated MIP-2 release in a dose-dependent manner. In vivo, pleural fluid MIP-2 levels in C57BL/6 mice injected intrapleurally with SLIGRL-NH2 (10 mg/kg) were significantly higher than in mice injected with LSIGRL-NH2 or PBS (2,710 ± 165 vs. 880 ± 357 vs. 88 ± 46 pg/ml, respectively; P < 0.001). Pleural fluid neutrophil counts were higher in SLIGRL-NH2 group than in the LSIGRL-NH2 and PBS groups (by 40- and 26-fold, respectively; P < 0.05). This study establishes that activation of mesothelial cell PAR2 potently induces the release of inflammatory cytokines in vitro and neutrophil recruitment into the pleural cavity in vivo.

scholarly journals MiR-29b suppresses peritoneal metastases through inhibition of the mesothelial-mesenchymal transition (MMT) of human peritoneal mesothelial cells

2021 ◽  
Author(s):  
Yuki Kimura ◽  
Hideyuki Ohzawa ◽  
Hideyo Miyato ◽  
Yuki Kaneko ◽  
Kazuya Takahashi ◽  
...  
Keyword(s):  
Morphological Changes ◽  
Negative Control ◽  
Mesothelial Cells ◽  
Peritoneal Metastases ◽  
Dependent Manner ◽  
Mesenchymal Transition ◽  
Peritoneal Mesothelial Cells ◽  
Human Peritoneal Mesothelial Cells ◽  
And Migration

Abstract Background: Peritoneal dissemination is a major metastatic pathway for gastrointestinal and ovarian malignancies. The miR-29b family is downregulated in peritoneal fluids in patients with peritoneal metastases (PM). We examined the effect of miR-29b on mesothelial cells (MC) which play critical a role in the development of PM through mesothelial-mesenchymal transition (MMT). Methods: Human peritoneal mesothelial cells (HPMCs) were isolated from surgically resected omental tissue and MMT induced by stimulation with 10 ng/ml TGF-b1. MiR-29b mimics and negative control miR were transfected by lipofection using RNAiMAX and the effects on the MMT evaluated in vitro. Results: HPMC produced substantial amounts of miR-29b which was markedly inhibited by TGF-b1. TGF-b1 stimulation of HPMC induced morphological changes with decreased expression of E-cadherin and calretinin, and increased expression of vimentin and fibronectin. TGF-b1 also enhanced proliferation and migration of HPMC as well as adhesion of tumor cells in a fibronectin dependent manner. However, all events were strongly abrogated by simultaneous transfection of miR-29b. Conclusion: MiR-29b inhibits TGF-b1 induced MMT and replacement of miR-29b in the peritoneal cavity might be effective to prevent development of PM partly through the effects on MC.

Insulin Stimulates the Activity of Na+/K+-Atpase in Human Peritoneal Mesothelial Cells

1997 ◽  
Vol 17 (2) ◽  
pp. 186-193 ◽  
Author(s):  
Janusz Witowski ◽  
Andrzej Breborowicz ◽  
Nicholas Topley ◽  
Leo Martis ◽  
Jan Knapowski ◽  
...  
Keyword(s):  
Atpase Activity ◽  
Mesothelial Cells ◽  
Pcr Analysis ◽  
Dependent Manner ◽  
Rt Pcr ◽  
Sodium Influx ◽  
Atpase Subunit ◽  
Peritoneal Mesothelial Cells ◽  
Human Peritoneal Mesothelial Cells ◽  
Dose Dependent

Objective To assess the effect of insulin on the Na+/ K+-ATPase expression and activity in human peritoneal mesothelial cells (HPMC). Methods HPMC were isolated from the omental tissue of non-uremic patients, grown to confluence and rendered quiescent by serum deprivation for 24 hours. The activity of Na+/K+-ATPase was determined by measuring the ouabain-sensitive86Rb uptake. To assess whether the effect of insulin was related to changes in [Na+]i the sodium influx was measured with 22Na and the activity of Na+/K+ -A TPase was assessed in the presence of amiloride. Expression of Na+/K+ -A TPaseα1’ α2 and β1-subunit mRNAs was determined by RT/PCR. Results Exposure of HPMC to insulin resulted in a time and dose-dependent increase in the Na+/K+-ATPase activity. After 60 minutes the ouabain-sensitive 86Rb up take (cpm/104 cells) was increased from 6650±796 in control cells to 9763±1212 in HPMC exposed to 100 mU/ mL insulin (1.5-fold increase; n=4, P<0.05). In addition, incubation of HPMC with 100 mU/mL insulin resulted in a time-dependent increase in the 22Na influx. Pre-exposure of HPMC to 1 mM amiloride reduced the activity of Na+/K+-A TPase but did not block the stimulatory effect of insulin. RT/PCR analysis revealed that HPMC constitutively expressed α1 and β1-subunit mRNAs while the α2-subunit mRNA was barely detectable. Exposure of HPMC to insulin for up to 24 hours was not associated with any changes in the expression of either α1’ α2 or B1-subunit. Conclusion Insulin stimulates the Na+/K+-ATPase activity in HPMC in a time and dose-dependent manner. This effect appears to mediated by an increase in [Na+]i and is not related to alterations in Na+/K+-ATPase subunit mRNAs expression.

Mesothelial Cells

2007 ◽  
Vol 27 (2_suppl) ◽  
pp. 110-115 ◽  
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.

Glucose but not N-Acetylglucosamine Accelerates in vitro Senescence of Human Peritoneal Mesothelial Cells

2011 ◽  
Vol 34 (6) ◽  
pp. 489-494 ◽  
Author(s):  
Marta Ciszewicz ◽  
George Wu ◽  
Paul Tam ◽  
Alicja Połubinska ◽  
Andrzej Bręborowicz
Keyword(s):  
Mesothelial Cells ◽  
Peritoneal Mesothelial Cells ◽  
Human Peritoneal Mesothelial Cells

Catabolism of Newly Synthesized Decorin in vitro by Human Peritoneal Mesothelial Cells

2004 ◽  
Vol 24 (2) ◽  
pp. 147-155 ◽  
Author(s):  
Susan Yung ◽  
Heinz Hausser ◽  
Gareth Thomas ◽  
Liliana Schaefer ◽  
Hans Kresse ◽  
...  
Keyword(s):  
Dermatan Sulfate ◽  
Specific Activity ◽  
Cell Layer ◽  
Mesothelial Cells ◽  
Matrix Assembly ◽  
Peritoneal Mesothelial Cells ◽  
Intracellular Degradation ◽  
Human Peritoneal Mesothelial Cells ◽  
Cell Medium

Objective Previous studies have shown that decorin and biglycan account for over 70% of the proteoglycans (PGs) synthesized by human peritoneal mesothelial cells (HPMCs). Since these PGs are involved in the control of cell growth, cell differentiation, and matrix assembly, we investigated their turnover in cultured HPMCs. Methods Confluent HPMCs were metabolically labeled with [35S]-sulfate and the labeled products isolated from the cell medium and the cell layer characterized by sensitivity to bacterial eliminases. Experiments were undertaken with exogenous labeled decorin, and its metabolic state was studied. Results In a 24-hour labeling period, 75% of the newly synthesized chondroitin sulfate/dermatan sulfate (CS/DS) PGs appeared in the culture medium, the majority of which (90%) was decorin. In the cell layer, protein-free glycosaminoglycan (GAG) chains accounted for 21% of the total CS/DS at 24 hours and exhibited constant specific activity at 12 – 16 hours. The latter material was turned over with a half-life of approximately 2.5 hours. Exogenous decorin underwent receptor-mediated endocytosis and subsequent intracellular degradation. Uptake but not degradation could be inhibited by heparin. Conclusions HPMCs are distinguished by a rapid turnover of decorin. A characteristic metabolic feature is the existence of a large intracellular pool of protein-free DS-GAGs. Understanding the control of decorin turnover in HPMCs might lead to delineation of its potential role in both the physiology and pathophysiology of the membrane in PD patients.

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