Ex vivo normothermic perfusion of isolated segmental porcine bowel: a novel functional model of the small intestine

Author:

Hamed M O1,Barlow A D1,Dolezalova N1,Khosla S2,Sagar A2,Gribble F M3,Davies S3,Murphy M P4,Hosgood S A1,Nicholson M L1,Saeb-Parsy K1

Affiliation:

1. Department of Surgery, University of Cambridge, and NIHR Cambridge Biomedical Research Campus, Cambridge, UK

2. Wellcome Trust – MRC Institute of Metabolic Science, University of Cambridge, Cambridge, UK

3. Department of Histopathology, University of Cambridge, Cambridge, UK

4. MRC Mitochondrial Biology Unit, Cambridge, UK

Abstract

Abstract Background There is an unmet need for suitable ex vivo large animal models in experimental gastroenterology and intestinal transplantation. This study details a reliable and effective technique for ex vivo normothermic perfusion (EVNP) of segmental porcine small intestine. Methods Segments of small intestine, 1.5–3.0 m in length, were retrieved from terminally anaesthetized pigs. After a period of cold ischaemia, EVNP was performed for 2 h at 37°C with a mean pressure of 80 mmHg using oxygenated autologous blood diluted with Ringer’s solution. The duration of EVNP was extended to 4 h for a second set of experiments in which two segments of proximal to mid-ileum (1.5–3.0 m) were retrieved from each animal and reperfused with whole blood (control) or leucocyte-depleted blood to examine the impact of leucocyte depletion on reperfusion injury. Results After a mean cold ischaemia time of 5 h and 20 min, EVNP was performed in an initial group of four pigs. In the second set of experiments, five pigs were used in each group. In all experiments bowel segments were well perfused and exhibited peristalsis during EVNP. Venous glucose levels significantly increased following luminal glucose stimulation (mean(s.e.m.) basal level 1.8(0.6) mmol/l versus peak 15.5(5.8) mmol/l; P < 0.001) and glucagon-like peptide 1 (GLP-1) levels increased in all experiments, demonstrating intact absorptive and secretory intestinal functions. There were no significant differences between control and leucocyte-depleted animals regarding blood flow, venous glucose, GLP-1 levels or histopathology at the end of 4 h of EVNP. Conclusions This novel model is suitable for the investigation of gastrointestinal physiology, pathology and ischaemia reperfusion injury, along with evaluation of potential therapeutic interventions.

Publisher

Oxford University Press (OUP)

Subject

General Medicine

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