Peritoneal Mesothelial Cell Culture and Biology

The peritoneal mesothelium is composed of an extensive monolayer of mesothelial cells that lines the body's serous cavity and internal organs and was previously thought to act principally as a protective nonadhesive lubricating surface to facilitate intracoelomic movement. With the introduction of peritoneal dialysis over three decades ago, there has been much interest in the cell biology of peritoneal mesothelial cells. Independent studies have highlighted specific properties of the peritoneal mesothelial cell, including antigen presentation, regenerative properties, clearance of fibrin; synthesis of cytokines, growth factors, and matrix proteins; and secretion of lubricants to protect the tissue from abrasion, adhesion, infection, and tumor dissemination. It is now evident that the mesothelium is not merely a passive membrane but, rather, a dynamic membrane that contributes substantially to the structural, functional, and homeostatic properties of the peritoneum. Since peritoneal mesothelial cells in culture possess immunohistochemical markers identical to mesothelial stem cells, the culture of mesothelial cells offers researchers an essential tool to assess their morphologic, structural, and functional properties. This review will discuss current procedures to isolate peritoneal mesothelial cells from human omental specimens, animal sources, and spent dialysate. Furthermore, the functional and morphologic properties of mesothelial cells are discussed, together with the potential use of mesothelial cell culture in research and clinical applications.

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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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TMIGD1 Inhibited Abdominal Adhesion Formation by Alleviating Oxidative Stress in the Mitochondria of Peritoneal Mesothelial Cells

Oxidative Medicine and Cellular Longevity ◽

10.1155/2021/9993704 ◽

2021 ◽

Vol 2021 ◽

pp. 1-16

Author(s):

Yunhua Wu ◽

Enmeng Li ◽

Zijun Wang ◽

Tianli Shen ◽

Cong Shen ◽

...

Keyword(s):

Oxidative Stress ◽

Mitochondrial Function ◽

Adhesion Formation ◽

Mesothelial Cell ◽

Mesothelial Cells ◽

Stress Injury ◽

Peritoneal Mesothelial Cells ◽

Abdominal Adhesion ◽

Adhesion Tissue ◽

Peritoneal Mesothelial Cell

Background. Postoperative abdominal adhesion remains one of the frequent complications after abdominal surgery and lacks effective intervention. Peritoneal mesothelial cell injury and healing play crucial roles in the process of adhesion formation, and identifying this mechanism might provide new insight into possible new therapeutic strategies for this disease. Transmembrane and immunoglobulin domain-containing 1 (TMIGD1) has been proven to protect renal epithelial cells from injury induced by oxidative stress and has also been identified as a novel adhesion molecule. Here, we investigated the role of TMIGD1 and its possible mechanism in adhesion formation. Materials and Methods. Immunohistochemistry (IHC), qPCR, and immunofluorescence (IHF) were used to detect the expression of TMIGD1. The grade and tenacity score of adhesion were used to evaluate the adhesion formation conditions. A TMIGD1-overexpressing HMrSV5 cell line was established. MTT assay, Western blotting, Annexin V apoptosis analysis, and CK19 staining were used to measure mesothelial cell viability, apoptosis, and completeness. ROS and MDA detection were used to measure mesothelial cell oxidative stress levels. JC-1 staining, IHF, and transmission electron microscopy were performed to assess mitochondrial function. Scratch-wound and adhesion assays were used to evaluate the adhesion ability of mesothelial cells. Results. First, we showed that TMIGD1 was decreased in mouse abdominal adhesion tissue and peritoneal mesothelial cells. Second, TMIGD1 overexpression inhibited adhesion formation. Third, TMIGD1 overexpression protected mesothelial cells from hydrogen peroxide- (H2O2-) induced oxidative stress injury. Fourth, TMIGD1 overexpression alleviated oxidative stress by protecting the mitochondrial function of mesothelial cells. In addition, TMIGD1 overexpression enhanced mesothelial cell adhesion. Conclusion. Our findings suggest that TMIGD1 protects mesothelial cells from oxidative stress injury by protecting their mitochondrial function, which is decreased in regular abdominal adhesion tissue. In addition, TMIGD1 enhances peritoneal mesothelial cell adhesion to promote healing.

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Acetylation of HMGB1 by JNK1 Signaling Promotes LPS-Induced Peritoneal Mesothelial Cells Apoptosis

BioMed Research International ◽

10.1155/2018/2649585 ◽

2018 ◽

Vol 2018 ◽

pp. 1-12 ◽

Cited By ~ 1

Author(s):

Shirong Cao ◽

Shu Li ◽

Yating Wang ◽

Jiani Shen ◽

Yi Zhou ◽

...

Keyword(s):

Posttranslational Modifications ◽

Cell Injury ◽

Mesothelial Cell ◽

High Mobility ◽

Mesothelial Cells ◽

Wild Type ◽

Mice Model ◽

Peritoneal Mesothelial Cells ◽

Visceral Peritoneum ◽

Peritoneal Mesothelial Cell

Increased high mobility group box 1 (HMGB1) in dialysis effluence is associated with the presence of peritoneal dialysis-related peritonitis in patients and peritoneal dysfunction in acute peritonitis mice model, but it remains unclear whether HMGB1 is involved in peritoneal mesothelial cell injury and functions via molecular posttranslational modifications by acetylation in this process. Here we first showed correlation between HMGB1 acetylation level in dialysis effluence of patients and occurrence of Gram-negative peritonitis. The increased level of acetylated HMGB1 was similarly observed under the lipopolysaccharides (LPS) treatment in both human peritoneal mesothelial cell line (HMrSV5) and mice visceral peritoneum tissue. Overexpression of wild-type, but not hypoacetylation mutant of HMGB1, enhanced LPS-induced apoptosis in HMrSV5 cells, which was accompanied by elevated protein levels of BAX and cleaved-caspase 3 compared to the control. Pretreatment of HMrSV5 cell with JNK inhibitor attenuated LPS-induced HMGB1 acetylation. Consistently, primary peritoneal mesothelial cells from Jnk1-/- mice showed a lower protein contents of acetylated HMGB1, fewer apoptosis, and decreased protein expression of BAX and cleaved-caspase3 after LPS exposure, as compared to those from wild-type mice. In conclusion, our data demonstrated HMGB1 promotes LPS-induced peritoneal mesothelial cells apoptosis, which is associated with JNK1-mediated upregulation of HMGB1 acetylation.

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Isolation and Propagation in Vitro of Peritoneal Mesothelial Cells

Peritoneal Dialysis International ◽

10.1177/089686088900900423 ◽

1989 ◽

Vol 9 (4) ◽

pp. 341-347 ◽

Cited By ~ 41

Author(s):

J. Thomas Hjelle ◽

Barbara T. Golinska ◽

Diane C. Waters ◽

Kevin R. Steidley ◽

David R. McCarroll ◽

...

Keyword(s):

Peritoneal Dialysis ◽

Cell Biology ◽

Life Support ◽

Cultured Cells ◽

Mesothelial Cell ◽

Experimental System ◽

Mesothelial Cells ◽

Growth Media ◽

Peritoneal Mesothelial Cells

Mesothelial cells lining the peritoneal cavity are the primary site of molecular exchange during peritoneal dialysis, a life support system for over 50 000 patients worldwide. In this study, techniques are described for the isolation and propagation in culture of peritoneal mesothelial cells from rats and rabbits. For comparison, mesothelial cells were also obtained from the serosal surface of human colonic tissue. By electron microscopy the cultured cells were found to exhibit microvilli, a well developed endoplasmic reticulum and golgi apparatus, micropinocytotic vesicles, and lipid-filled intracellular vesicles. Immunochemical probes revealed the expression by these cells in vitro of cytokeratin, fibronectin, vimentin, and keratin, but not von Willebrand factor. Mesothelial cells from rat, rabbit, and human exhibited contact inhibition, but differences in growth rates and dependence on supplements to the growth media. This work provides a multispecies comparison of the behavior of mesothelial cells in vitro for the purpose of developing an experimental system for the study of mesothelial cell biology and the role of these cells in peritoneal dialysis.

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Effect of High Glucose on Peritoneal Mesothelial Cell Biology

Peritoneal Dialysis International ◽

10.1177/089686080002002s04 ◽

2000 ◽

Vol 20 (2_suppl) ◽

pp. 15-18 ◽

Cited By ~ 43

Author(s):

Hunjoo Ha ◽

Hi Bahl Lee

Keyword(s):

Protein Synthesis ◽

Transforming Growth Factor Beta ◽

High Glucose ◽

Cell Biology ◽

Transforming Growth Factor ◽

Mesothelial Cell ◽

Continuous Exposure ◽

Messenger Rnas ◽

Peritoneal Fibrosis ◽

Peritoneal Mesothelial Cell

Objective This study reviews evidence that implicates high glucose (HG) in the pathogenesis of peritoneal fibrosis and proposes mechanisms potentially involved in the HG-induced peritoneal fibrosis that is observed in long-term peritoneal dialysis (PD) patients. Design Selected Western literature is reviewed, examining the effect of HG on rat or human peritoneal mesothelial cell (HPMC) biology with particular reference to extracellular matrix (ECM) gene expression and protein synthesis. Results HG up-regulated the expression of monocyte chemotactic peptide–1 (MCP-1), transforming growth factor beta 1 (TGFβ1), and fibronectin messenger RNAs (mRNAs) and proteins. These HG-induced up-regulations were effectively blocked by the inhibition of protein kinase C (PKC). In addition, cytosolic reactive oxygen species (ROS) rapidly increased in HPMC cultured under HG, and treatment with antioxidant effectively inhibited HG-induced fibronectin protein synthesis by HPMC. Conclusion Continuous exposure of the peritoneal membrane to HG may induce changes in HPMC biology, leading to excessive deposition of ECM and peritoneal injury. HG-induced activation of diacylglycerol PKC (DAG–PKC) plays a major role in up-regulation of MCP-1, TGFβ1, and fibronectin synthesis by HPMC cultured under HG. In addition, ROS, recently recognized as signalling molecules, are rapidly generated in HPMC as a result of increased glucose metabolism and may prove to be an important mediator of HG-induced peritoneal injury.

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Electrophysiology and Glucose Transport of Human Peritoneal Mesothelial Cells: Implications for Peritoneal Dialysis

Peritoneal Dialysis International ◽

10.1177/089686080102100202 ◽

2001 ◽

Vol 21 (2) ◽

pp. 115-122 ◽

Cited By ~ 7

Author(s):

Fu Keung Li ◽

Chi Ho To ◽

Jack Kok Hung Leung ◽

Tak Mao Chan ◽

Ka Neng Lai

Keyword(s):

Glucose Transport ◽

Ussing Chamber ◽

Mesothelial Cells ◽

Resting Potential ◽

Peritoneal Mesothelial Cells ◽

Ussing Chambers ◽

Serial Sampling ◽

Human Peritoneal Mesothelial Cells ◽

Peritoneal Mesothelium ◽

Paracellular Shunt

Objective To elucidate ionic and glucose transport across human peritoneal mesothelium, we utilized an Ussing chamber setup and studied the electrophysiological characteristics and tissue permeabilities of human peritoneal mesothelial cells (HPMC) to l- and d-glucose. Methods Human mesothelial cells were grown on polyester filters (snapwell; Costar, Cambridge, MA, U.S.A.) that, upon confluence, were fitted into Ussing chambers. Transmesothelial resistance and resting potential were determined using electro-physiological techniques. Radiolabeled glucose was added to one side of the chamber and the permeabilities determined by serial sampling in the receptive compartment. Results The transmesothelial potential and resistance were 0.54 ± 0.07 mV (apical positive) and 20.4 ± 3.2 Ω·cm2 respectively (mean ± SEM, n = 36). The course of overall transfer of d- and l-glucose was examined using l-glucose as a positive diffusion-plus-leak marker. The permeabilities of HPMC to d-glucose were 3.00 ± 0.26 cm/sec (apical-to-basolateral) and 3.25 ± 0.27 cm/sec (basolateral-to-apical) [ n = 6 experiments, p = not significant (NS)], which were not different from those of l-glucose: 3.00 ± 0.30 cm/sec (apical-to-basolateral) and 2.71 ± 0.24 (basolateral-to-apical) ( n = 6 experiments, p = NS). Conclusions The transepithelial resistance of HPMC is low and the ionic gradient, although it exists, is small and inconsequential. Passive paracellular flow accounts for the majority of transmesothelial glucose transport. The existence of a large paracellular shunt precludes the mesothelial membrane as a clinically relevant osmotic barrier.

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Gastric cancer derived exosomes induce peritoneal mesothelial cell EMT through TGF-β1/Smads pathway to promote peritoneal metastasis

10.1101/2021.07.23.453534 ◽

2021 ◽

Author(s):

Jungang Dong ◽

Zhongbo Zhu ◽

Guoning Cui ◽

Zhixuan Zhang ◽

Juan Yue ◽

...

Keyword(s):

Gastric Cancer ◽

Peritoneal Metastasis ◽

Epithelial Mesenchymal Transition ◽

Critical Role ◽

Mesothelial Cell ◽

Mesothelial Cells ◽

Mesenchymal Transition ◽

Peritoneal Mesothelial Cells ◽

Metastatic Microenvironment ◽

Tgf Β1

Epithelial-mesenchymal transition (EMT) plays an important role in peritoneal metastasis of Gastric cancer (GC). Tumor exosomes can mediate tumor directed metastasis, and TGF-β1 is an important factor in inducing tumor Epithelial mesenchymal transition. However, it is not clear whether GC derived exosomes can induce peritoneal mesothelial cells through the TGF-β1/ Smads pathway and the effect of injured peritoneal mesothelial cells on the biological characteristics of GC cells. In this study, we demonstrated that GC-derived exosomes can activate the TGF-β1/Smads pathway in peritoneal mesothelial cells and induce the corresponding EMT process, and that the injured peritoneal mesothelial cells can improve the migration and adhesion of GC cells. Taken together, these data further support the critical role of exosomes in the remodeling of the pre-metastatic microenvironment.

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

Cell Transplantation ◽

10.1177/096368979400300107 ◽

1994 ◽

Vol 3 (1) ◽

pp. 41-48 ◽

Cited By ~ 3

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.

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miRNA589 Regulates Epithelial-Mesenchymal Transition in Human Peritoneal Mesothelial Cells

Journal of Biomedicine and Biotechnology ◽

10.1155/2012/673096 ◽

2012 ◽

Vol 2012 ◽

pp. 1-6 ◽

Cited By ~ 15

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.

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Human peritoneal mesothelial cells produce many cytokines (granulocyte colony-stimulating factor [CSF], granulocyte-monocyte-CSF, macrophage- CSF, interleukin-1 [IL-1], and IL-6) and are activated and stimulated to grow by IL-1

Blood ◽

10.1182/blood.v80.11.2835.2835 ◽

1992 ◽

Vol 80 (11) ◽

pp. 2835-2842 ◽

Cited By ~ 103

Author(s):

L Lanfrancone ◽

D Boraschi ◽

P Ghiara ◽

B Falini ◽

F Grignani ◽

...

Keyword(s):

Interleukin 1 ◽

Mesothelial Cell ◽

Mesothelial Cells ◽

Type I ◽

Colony Stimulating Factor ◽

Peritoneal Mesothelial Cells ◽

Gm Csf ◽

Regulatory Molecule ◽

Macrophage Colony ◽

Stimulating Factor

Abstract To investigate the role of peritoneal mesothelial cells in regulating hematopoiesis, as well as inflammation, healing, and tissue regeneration processes, long-term cultures of peritoneal mesothelial cells from human endocavitarian fluids were established. The purity of the cell population was assessed by morphologic and immunocytochemical criteria. Five peritoneal mesothelial cell cultures were analyzed for cytokine expression. Macrophage colony-stimulating factor (M-CSF), granulocyte-CSF (G-CSF), interleukin-1 alpha (IL-1 alpha), IL-1 beta, and IL-6 transcripts were constantly but variably detected throughout the culture period, while granulocyte-monocyte-CSF (GM-CSF) expression started as the cell culture aged. No IL-2, IL-3, IL-4, IL-5, or IL-7 transcripts were detected in the same samples. Corresponding cytokine activities were detected in the supernatants of the cultures. Peritoneal mesothelial cells proliferated after the addition of exogenous IL-1 beta or IL-1 alpha, whereas the addition of recombinant GM-CSF, G-CSF, M-CSF, or IL-6 failed to trigger proliferation. IL-1 receptor type I transcripts were detected in peritoneal mesothelial cells. Moreover, IL-1 was able to upregulate the expression of the genes that code for G-CSF, GM-CSF, IL-1 alpha, and IL-1 beta in these cells. These data indicate that peritoneal mesothelial cells produce many cytokines and suggest that IL-1 is a regulatory molecule for peritoneal mesothelial cells.

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