title: Precision-cut kidney slices (PCKS) to study development of renal fibrosis and efficacy of drug targeting ex vivo.
abstract: Renal fibrosis is a serious clinical problem resulting in the greatest need for renal replacement therapy. No adequate preventive or curative therapy is available that could be clinically used to target renal fibrosis specifically. The search for new efficacious treatment strategies is therefore warranted. Although in vitro models using homogeneous cell populations have contributed to the understanding of the pathogenetic mechanisms involved in renal fibrosis, these models poorly mimic the complex in vivo milieu. Therefore, we here evaluated a precision-cut kidney slice (PCKS) model as a new, multicellular ex vivo model to study the development of fibrosis and its prevention using anti-fibrotic compounds. Precision-cut slices (200-300 μm thickness) were prepared from healthy C57BL/6 mouse kidneys using a Krumdieck tissue slicer. To induce changes mimicking the fibrotic process, slices were incubated with TGFβ1 (5 ng/ml) for 48 h in the presence or absence of the anti-fibrotic cytokine IFNγ (1 µg/ml) or an IFNγ conjugate targeted to PDGFRβ (PPB-PEG-IFNγ). Following culture, tissue viability (ATP-content) and expression of α-SMA, fibronectin, collagen I and collagen III were determined using real-time PCR and immunohistochemistry. Slices remained viable up to 72 h of incubation, and no significant effects of TGFβ1 and IFNγ on viability were observed. TGFβ1 markedly increased α-SMA, fibronectin and collagen I mRNA and protein expression levels. IFNγ and PPB-PEG-IFNγ significantly reduced TGFβ1-induced fibronectin, collagen I and collagen III mRNA expression, which was confirmed by immunohistochemistry. The PKCS model is a novel tool to test the pathophysiology of fibrosis and to screen the efficacy of anti-fibrotic drugs ex vivo in a multicellular and pro-fibrotic milieu. A major advantage of the slice model is that it can be used not only for animal but also for (fibrotic) human kidney tissue.
authors.affiliation: Departments of Pathology and Medical Biology, Division of Pathology, University Medical Center Groningen, University of Groningen, Groningen, 9713 GZ, The Netherlands.
authors.forename: Fariba
authors.initials: F
authors.lastname: Poosti
authors.affiliation: Department of Pharmaceutical Technology and Biopharmacy, University of Groningen, Groningen, 9713 AV, The Netherlands.
authors.forename: Bao Tung
authors.initials: BT
authors.lastname: Pham
authors.affiliation: Department of Pharmaceutical Technology and Biopharmacy, University of Groningen, Groningen, 9713 AV, The Netherlands.
authors.forename: Dorenda
authors.initials: D
authors.lastname: Oosterhuis
authors.affiliation: Department of Pharmacokinetics, Toxicology and Targeting, University of Groningen, Groningen, 9713 AV, The Netherlands.
authors.forename: Klaas
authors.initials: K
authors.lastname: Poelstra
authors.affiliation: Departments of Pathology and Medical Biology, Division of Pathology, University Medical Center Groningen, University of Groningen, Groningen, 9713 GZ, The Netherlands.
authors.forename: Harry
authors.initials: H
authors.lastname: van Goor
authors.affiliation: Department of Pharmaceutical Technology and Biopharmacy, University of Groningen, Groningen, 9713 AV, The Netherlands.
authors.forename: Peter
authors.initials: P
authors.lastname: Olinga
authors.affiliation: Departments of Pathology and Medical Biology, Division of Pathology, University Medical Center Groningen, University of Groningen, Groningen, 9713 GZ, The Netherlands j.l.hillebrands@umcg.nl.
authors.forename: Jan-Luuk
authors.initials: JL
authors.lastname: Hillebrands
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D024043:Collagen Type II / Q000378:metabolism; D016503:Drug Delivery Systems; D005109:Extracellular Matrix / Q000187:drug effects / Q000378:metabolism; D005353:Fibronectins / Q000378:metabolism; D005355:Fibrosis; D016650:Fluorescein-5-isothiocyanate / Q000378:metabolism; D000949:Histocompatibility Antigens Class II / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007371:Interferon-gamma / Q000378:metabolism; D007668:Kidney / Q000187:drug effects / Q000473:pathology*; D008810:Mice, Inbred C57BL; D008954:Models, Biological; D009113:Muramidase / Q000378:metabolism; D011092:Polyethylene Glycols / Q000737:chemistry; D020797:Receptor, Platelet-Derived Growth Factor beta / Q000235:genetics / Q000378:metabolism; D013997:Time Factors; D014022:Tissue Survival / Q000187:drug effects; D053773:Transforming Growth Factor beta1 / Q000494:pharmacology; D016896:Treatment Outcome
chemical_list: D015415:Biomarkers; D024043:Collagen Type II; D005353:Fibronectins; D000949:Histocompatibility Antigens Class II; D053773:Transforming Growth Factor beta1; D011092:Polyethylene Glycols; D007371:Interferon-gamma; D020797:Receptor, Platelet-Derived Growth Factor beta; D009113:Muramidase; D016650:Fluorescein-5-isothiocyanate
keywords: Drug targeting; Fibrosis; Kidney; Tissue slices