Precision-Cut Kidney Slices as a Tool to Understand the Dynamics of Extracellular Matrix Remodeling in Renal Fibrosis

Abstract

The aim of this study was to set up an ex vivo model for renal interstitial fibrosis in order to investigate the extracellular matrix (ECM) turnover profile in the fibrotic kidney. We induced kidney fibrosis in fourteen 12-week-old male Sprague Dawley rats by unilateral ureteral obstruction (UUO) surgery of the right ureter. The left kidney (contralateral) was used as internal control. Six rats were sham operated and used as the control group. Rats were terminated two weeks after the surgery; the kidneys were excised and precision-cut kidney slices (PCKSs) were cultured for five days in serum-free medium. Markers of collagen type I formation (P1NP), collagen type I and III degradation (C1M and C3M), and α-smooth muscle actin (αSMA) were measured in the PCKS supernatants by enzyme-linked immunosorbent assay. P1NP, C1M, C3M, and α-SMA were increased up to 2- to 13-fold in supernatants of tissue slices from the UUO-ligated kidneys compared with the contralateral kidneys (P < 0.001) and with the kidneys of sham-operated animals (P < 0.0001). The markers could also reflect the level of fibrosis in different animals. The UUO PCKS ex vivo model provides a valuable translational tool for investigating the extracellular matrix remodeling associated with renal interstitial fibrosis.

Murine Precision-Cut Kidney Slices as an ex vivo Model to Evaluate the Role of Transforming Growth Factor-β1 Signaling in the Onset of Renal Fibrosis

“…Translational models of renal fibrosis to develop efficacious antifibrotic therapies are lacking (Stribos et al, 2016a ; Klinkhammer et al, 2017 ) and there is an urgent need to close the gap between in vivo studies and clinical trials. PCKS have been around for several decades for pharmacotoxicology studies and recently this ex vivo model has been implemented in the fibrosis research field (Poosti et al, 2015 ; Genovese et al, 2016 ; Zhang et al, 2016 ). Strengths of this promising technique are the preserved multicellular heterogeneity and organ architecture, the possibility to perform translational research using human PCKS and the potential to replace/reduce animal experiments.…”

Inhibition of tyrosine kinase receptor signaling attenuates fibrogenesis in an ex vivo model of human renal fibrosis

“…In vitro models lack cellular heterogeneity, and animal models have limited implications for human disease. To partly overcome these limitations, we used precision-cut kidney slices (PCKS) as an ex vivo model of renal fibrosis (13,34,43). PCKS replicate the organotypic…”

Selecting the right therapeutic target for kidney disease

“…Historically, PCKS have been used to study renal fibrosis, a very complex process that cannot be elucidated using in vitro systems. Two strategies can be used: slices can be taken from injured kidneys, for example, after unilateral ureteral obstruction ( Genovese et al, 2016 ) or slices can be taken from healthy kidneys and treated with pro-fibrotic stimuli such as TGF-β ( Stribos et al, 2017 ). An advantage of PCKS is that they can be cultured for 5–7 days, making them suitable for longitudinal studies and amenable to the types of analyses that are performed on kidneys isolated from mice.…”