Establishment and functional characterization of human peritoneal fibroblasts in culture

Jörres, Achim; Ludat, K; Lang, Jochen; Sander, Karoline; Gahl, Gerhard M.; Frei, Ulrich; DeJonge, K; Williams, Julie; Topley, Nicholas

doi:10.1681/asn.v7102192

Cited by 50 publications

(6 citation statements)

References 24 publications

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“…Taken together, acute PDF exposure activates molecular pathways that involve a significant number of proteins being players in more than one cellular process. Whereas the majority of these processes contribute to protection of cells under acute stressful conditions, some of them may be deleterious for the peritoneal membrane in response to chronic PDF exposure. – Data like ours might allow identifying novel candidates as biomarkers.…”

Section: Discussionmentioning

confidence: 88%

“…Whereas the majority of these processes contribute to protection of cells under acute stressful conditions, some of them may be deleterious for the peritoneal membrane in response to chronic PDF exposure. [48][49][50] Data like ours might allow identifying novel candidates as biomarkers.…”

Section: Discussionmentioning

confidence: 89%

“…Peritoneal mesothelial cells are known to produce cytokines like IL-1β, IL-6, IL-8 and TNF-R and to overexpress adhesion molecules when stimulated. [48][49][50] Besides the group of molecular chaperones, our data revealed involvement of novel players like cyclophilin A (PPIA), peroxiredoxin (PRDX1) and growth factor receptorbound protein 2 (GRB2). PPIA not only shows activity in protein folding as a peptidylprolyl cis-trans-isomerase, 51 but is also a member of the immunophilin family 52 and the receptor for the immunosuppressive drugs tacrolimus and cyclosporine.…”

Section: Discussionmentioning

confidence: 93%

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Stress Responses and Conditioning Effects in Mesothelial Cells Exposed to Peritoneal Dialysis Fluid

et al. 2009

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Renal replacement therapy by peritoneal dialysis is frequently complicated by technical failure. Peritoneal dialysis fluids (PDF) cause injury to the peritoneal mesothelial cell layer due to their cytotoxicity. As only isolated elements of the involved cellular processes have been studied before, we aimed at a global assessment of the mesothelial stress response to PDF. Following single or repeated exposure to PDF or control medium, proteomics and bioinformatics techniques were combined to study effects in mesothelial cells (MeT-5A). Protein expression was assessed by two-dimensional gel electrophoresis, and significantly altered spots were identified by MALDI-TOF MS and MS2 techniques. The lists of experimentally derived candidate proteins were expanded by a next neighbor approach and analyzed for significantly enriched biological processes. To address the problem of an unknown portion of false positive spots in 2DGE, only proteins showing significant p-values on both levels were further interpreted. Single PDF exposure resulted in reduction of biological processes in favor of reparative responses, including protein metabolism, modification and folding, with chaperones as a major subgroup. The observed biological processes triggered by this acute PDF exposure mainly contained functionally interwoven multitasking proteins contributing as well to cytoskeletal reorganization and defense mechanisms. Repeated PDF exposure resulted in attenuated protein regulation, reflecting inhibition of stress responses by high levels of preinduced chaperones. The identified proteins were less attributable to acute cellular injury but rather to specialized functions with a reduced number of involved multitasking proteins. This finding agrees well with the concept of conditioning effects and cytoprotection. In conclusion, this study describes the reprogrammed proteome of mesothelial cells during recovery from PDF exposure and adaption to repetitive stress. A broad stress response with a number of highly overlapping processes and multitasking proteins shifts toward a more specific response of only few less overlapping processes.

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Section: Discussionmentioning

confidence: 88%

Section: Discussionmentioning

confidence: 89%

Section: Discussionmentioning

confidence: 93%

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Stress Responses and Conditioning Effects in Mesothelial Cells Exposed to Peritoneal Dialysis Fluid

et al. 2009

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“…Their role is mainly associated with the development of peritoneal fibrosis induced by chronic inflammation [ 104 ]. However, human peritoneal fibroblasts (HPFBs) upon stimulation with macrophage or T lymphocyte-derived proinflammatory cytokines, such as IL-1β, TNFα and IFN-γ, secrete large quantities of interleukins and chemokines (CCL2, IL-8/CXCL8, IL-6, CXCL1, CCL5) [ 101 , 105 , 106 , 107 , 108 ]. Therefore, HBFBs control trans-peritoneal chemotactic gradients during peritonitis, crucial when MCs are damaged and exfoliated.…”

Section: Resident Peritoneal Cells: Activation Of Proinflammatory And...mentioning

confidence: 99%

Novel Aspects of the Immune Response Involved in the Peritoneal Damage in Chronic Kidney Disease Patients under Dialysis

Trionfetti

Marchant

González-Mateo

et al. 2023

IJMS

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Chronic kidney disease (CKD) incidence is growing worldwide, with a significant percentage of CKD patients reaching end-stage renal disease (ESRD) and requiring kidney replacement therapies (KRT). Peritoneal dialysis (PD) is a convenient KRT presenting benefices as home therapy. In PD patients, the peritoneum is chronically exposed to PD fluids containing supraphysiologic concentrations of glucose or other osmotic agents, leading to the activation of cellular and molecular processes of damage, including inflammation and fibrosis. Importantly, peritonitis episodes enhance peritoneum inflammation status and accelerate peritoneal injury. Here, we review the role of immune cells in the damage of the peritoneal membrane (PM) by repeated exposure to PD fluids during KRT as well as by bacterial or viral infections. We also discuss the anti-inflammatory properties of current clinical treatments of CKD patients in KRT and their potential effect on preserving PM integrity. Finally, given the current importance of coronavirus disease 2019 (COVID-19) disease, we also analyze here the implications of this disease in CKD and KRT.

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“…Isolation of human pleural fibroblasts (HPF). The enzymatic disaggregation of tissue to isolate fibroblasts was modified from Joerres et al [24]. Pleural specimens were handled under sterile conditions, washed three times in PBS and pieces approximately 2 mm 2 in size were incubated in 5 ml trypsin/EDTA/glucose (0.125%/0.01%/0.1 wt/vol in PBS) for 20 min at 37˚C an orbital shaker.…”

Section: Cell Culturementioning

confidence: 99%

Novel 3D organotypic co-culture model of pleura

et al. 2022

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Pleural mesothelial cells are the predominant cell type in the pleural cavity, but their role in the pathogenesis of pleural diseases needs to be further elucidated. 3D organotypic models are an encouraging approach for an in vivo understanding of molecular disease development. The aim of the present study was to develop a 3D organotypic model of the pleural mesothelium. Specimens of human pleura parietalis were obtained from patients undergoing surgery at the University Hospital Leipzig, Germany. 3D co-culture model of pleura was established from human pleural mesothelial cells and fibroblasts. The model was compared to human pleura tissue by phase-contrast and light microscopy, immunochemistry and -fluorescence as well as solute permeation test. Histological assessment of the 3D co-culture model displayed the presence of both cell types mimicking the morphology of the human pleura. Vimentin and Cytokeratin, PHD1 showed a similar expression pattern in pleural biopsies and 3D model. Expression of Ki-67 indicates the presence of proliferating cells. Tight junctional marker ZO-1 was found localized at contact zones between mesothelial cells. Each of these markers were expressed in both the 3D co-culture model and human biopsies. Permeability of 3D organotypic co-culture model of pleura was found to be higher for 70 kDa-Dextran and no significant difference was seen in the permeability for small dextran (4 kDa). In summary, the presented 3D organoid of pleura functions as a robust assay for pleural research serving as a precise reproduction of the in vivo morphology and microenvironment.

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Establishment and functional characterization of human peritoneal fibroblasts in culture

Cited by 50 publications

References 24 publications

Stress Responses and Conditioning Effects in Mesothelial Cells Exposed to Peritoneal Dialysis Fluid

Stress Responses and Conditioning Effects in Mesothelial Cells Exposed to Peritoneal Dialysis Fluid

Novel Aspects of the Immune Response Involved in the Peritoneal Damage in Chronic Kidney Disease Patients under Dialysis

Novel 3D organotypic co-culture model of pleura

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