8. Porcine precision-cut kidney slices as an ex vivo model to evaluate the effect of a ketogenic diet on mitochondrial function

L.A. van Furth, H. Maassen, L.H. Venema, P. Olinga, H.G.D. Leuvenink

Wednesday 4 march 2020

0:00 - 0:00h at Toon Hermans Foyer

Parallel session: Postersessie 2 – Basale abstracts

Background: Kidneys derived from Donation after Circulatory Death (DCD) donors suffer from ischemic injury. When reperfusion occurs, the formation of Reactive Oxygen Species (ROS) contributes to the development of ischemia reperfusion injury (IRI). The major site of production and the major target of ROS are the mitochondria. Preoperative dietary restriction and fasting are known to be protective against renal IRI. During fasting or a ketogenic diet, the metabolic substrate switches from glucose to fatty acids, which then becomes the main source of adenosine triphosphate (ATP) production. Precision-cut kidney slices (PCKS) is an ex vivo model to evaluate intra- and extracellular mechanisms. Here, we aim to evaluate the effect of a ketogenic diet on mitochondrial function using PCKS.

Methods: Porcine kidneys were obtained at a local abattoir and prepared for hypothermic machine perfusion (HMP). After 30 min of WIT, kidneys were flushed and placed on HMP and preserved for 3 hours with UW-CS. Thereafter PCKS were made and incubated at 37ºC with 80% oxygen. The incubation medium of the control group consisted RPMI 1640 medium, which contains 2 mg/mL glucose. The experimental groups consisted RPMI 1640 medium without glucose, supplemented with 2 mg/mL (±)-sodium 3-hydroxybutyrate (BHB) or 1,5 mg/mL SMOFlipids®. Directly after slicing and after 24, 48 or 72 hours of incubation, tissue and medium samples were taken to analyse for different mitochondrial, oxidative stress and injury markers.

Results: After 72 hours of incubation in ketogenic culture medium all slices are vital. Improved mitochondrial function in both the experimental groups are observed compared to the control group. More in-depth analysis are ongoing.

Conclusions: A ketogenic culture environment seems to conserve PCKS by stabilizing mitochondria thereby protecting against IRI.