Journal Article

Precision-cut kidney slices (PCKS) to study development of renal fibrosis and efficacy of drug targeting ex vivo

Oosterhuis, Dorenda, van Goor, Harry, Poelstra, Klaas, Pham, Bao Tung, Olinga, Peter, Hillebrands, Jan-Luuk, Poosti, Fariba

2015

Overview

Renal fibrosis is a serious clinical problem forming the utmost cause of need for renal replacement therapy. No adequate preventive or curative therapy is available that can be clinically used to specifically target renal fibrosis. 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, here we evaluated a precision-cut kidney slice (PCKS) model as a new, multicellular ex vivo model to study development of fibrosis and the prevention thereof 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 hours in the presence or absence of the anti-fibrotic cytokine IFNγ (1 µg/ml) or an IFNγ conjugate which is targeted to the 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 hours 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. Major advantage of the slice model is that it can be used not only for animal but also for (fibrotic) human kidney tissue.

• Publisher: The Company of Biologists Ltd,The Company of Biologists
• Subject: Animals / Biological activity / Biomarkers - metabolism / Biopsy / Cell culture / Collagen Type II - metabolism / Cytokines / Drug Delivery Systems / Drug targeting / Experiments / Extracellular matrix / Extracellular Matrix - drug effects / Extracellular Matrix - metabolism / Fibroblasts / Fibronectins - metabolism / Fibrosis / Fluorescein-5-isothiocyanate - metabolism / Histocompatibility Antigens Class II - genetics / Histocompatibility Antigens Class II - metabolism / Humans / Interferon-gamma - metabolism / Kidney / Kidney - drug effects / Kidney - pathology / Kidney diseases / Mice, Inbred C57BL / Models, Biological / Muramidase - metabolism / Pathophysiology / Polyethylene Glycols - chemistry / Proteins / Receptor, Platelet-Derived Growth Factor beta - genetics / Receptor, Platelet-Derived Growth Factor beta - metabolism / Time Factors / Tissue slices / Tissue Survival - drug effects / Transforming Growth Factor beta1 - pharmacology / Transforming growth factor-b1 / Treatment Outcome