Exploring Porcine Precision-Cut Kidney Slices as a Model for Transplant-Related Ischemia-Reperfusion Injury

Abstract

Marginal donor kidneys are more likely to develop ischemia-reperfusion injury (IRI), resulting in inferior long-term outcomes. Perfusion techniques are used to attenuate IRI and improve graft quality. However, machine perfusion is still in its infancy, and more research is required for optimal conditions and potential repairing therapies. Experimental machine perfusion using porcine kidneys is a great way to investigate transplant-related IRI, but these experiments are costly and time-consuming. Therefore, an intermediate model to study IRI would be of great value. We developed a precision-cut kidney slice (PCKS) model that resembles ischemia-reperfusion and provides opportunities for studying multiple interventions simultaneously. Porcine kidneys were procured from a local slaughterhouse, exposed to 30 min of warm ischemia, and cold preserved. Subsequently, PCKS were prepared and incubated under various conditions. Adenosine triphosphate (ATP) levels and histological tissue integrity were assessed for renal viability and injury. Slicing did not influence tissue viability, and PCKS remained viable up to 72 h incubation with significantly increased ATP levels. Hypothermic and normothermic incubation led to significantly higher ATP levels than baseline. William’s medium E supplemented with Ciprofloxacin (and Amphotericin-B) provided the most beneficial condition for incubation of porcine PCKS. The porcine PCKS model can be used for studying transplant IRI.

Comparative analysis of kidney transplantation modeled using precision-cut kidney slices and kidney transplantation in pigs

“…Since severe ischemia morphologically resembles autolysis, this could have distorted the findings and led to misclassification of some ischemic features as autolysis. Presence or absence of autolysis was not reported in a previous study on porcine precision-cut kidney slices (14), and autolysis could potentially be decreased with future improvements in the PCKS methodology. However, the feature of autolysis of this model could potentially serve as a starting point to study the rapid degradation of some cytoskeletal proteins reported in kidney transplants from donors with brain death compared with transplants from living donors (28).…”

“…The presented study has several strengths. It is to our knowledge the first study to systematically compare a PCKS approach, an increasingly used methodology in kidney research (10)(11)(12)(13)(14), with a real disease model for kidney transplantation. We used in-depth morphological grading, allowing a detailed understanding of the morphological landscape throughout the study.…”

“…In recent years, several groups have examined precision-cut kidney slices (PCKS) of different species to model kidney injury, including endoplasmatic reticulum stress, fibrosis and antifibrotic drug evaluation (10–13). One experimental study used PCKS to determine the effect of different preservation solutions, temperature or cold ischemia on porcine kidney tissue viability (14). However, at present the potential usefulness of PCKS as a model to study kidney ischemia-reperfusion injury as the very early step of transplantation is unclear and more data are needed to evaluate how closely an ex vivo PCKS method resembles in vivo models, and how closely post-ischemic changes in PCKS mimic the pathophysiology of human kidney transplantation.…”

Targeted delivery of galunisertib using machine perfusion reduces fibrogenesis in an integrated ex vivo renal transplant and fibrogenesis model

“…Nevertheless, experimental normothermic machine perfusion models are commonly used platforms for optimizing the resuscitation technique itself, and ex viv o renal function accurately reflects the condition of the kidney (Venema et al, 2019, 2021; Weissenbacher et al, 2019). In contrast to renal function, we were able to assess renal mitochondrial activity for an additional 48 h using the well‐established technique of precision‐cut kidney slices (Bigaeva, Stribos, et al, 2020; van Furth et al, 2022; Jensen et al, 2019; van Leeuwen et al, 2022; Stribos et al, 2016; Tingskov et al, 2021; Vaughan et al, 2022). Kidneys are well‐known consumers of oxygen to facilitate oxidative phosphorylation for the active reabsorption of sodium in the tubular cells (Balaban, 1990; Lassen et al, 1961; Singh et al, 2013).…”

Modelling Acid-Induced Lung Damage in Precision-Cut Lung Slices: An Ex Vivo Animal Model

“…Anatomical structures and cell interactions of the lung are maintained. This technique has already proven its success in the fields of physiology, pharmacology, and toxicology [26][27][28]. In addition, mice have been accepted as a relatively accurate representation in the context of acute lung injury [26,29].…”