The perfused porcine pancreas as a model for testing organ protective solutions

Abstract

The present study was designed to establish an in vitro perfused porcine pancreas preparation as a model for testing the effect of organ protective solutions on stimulated pancreatic endocrine and exocrine secretion. The pancreas was prepared and perfused for 10 min with Euro Collins solution, thereafter it was stored in the cold (4 degrees C) for various times. After 3-h and 6-h ischemia pancreatic insulin release in response to glucose was not significantly affected. After 12-h ischemia reduced pancreatic insulin secretin, increased perfusion pressure, and increased amylase and lipase release indicated pancreatic damage. Complete pancreatic dysfunction was seen after 24-h and 48-h ischemia with massive increase in perfusion pressure and low insulin secretion which did not follow a glucose-dependent release pattern, while amylase and lipase concentrations in the perfusion medium increased. Stimulated exocrine pancreatic secretion was significantly decreased already after 3-h ischemia and completely lost after 12 h.

Preservation of the porcine pancreas with HTK and Euro‐Collins solution: studies in a reperfusion system

“…The present study evaluates the effect of the HTK solution on the preservation of the porcine pancreas using an in zritro model under controlled and defined conditions. This model was previously shown to react physiologically to both endocrine and exocrine stimuli and allows the testing of multiple parameters after ischaemia [7]. It therefore allows a detailed testing of the effect of a protective solution on pancreatic organ preservation.…”

“…The reperfusion chamber was placed in a water bath at 38'C. The pancreas was perfused in an arteriovenous one-line system with an oxygenated Krebs-Ringer bicarbonate buffer after addition of homologous erythrocytes to obtain a haematocrit of 10% [6,7]. The arterial perfusion pressure (Statham P 23 Db, Monitor SP 1405; Gould, Bilthoven, The Netherlands) was continuously monitored and was kept at 50 mmHg during the reperfusion period by adjusting the pump flow rate.…”

“…Pancreatic insulin release per minute was quantified in the perfusion medium as previously described [7]. Insulin was determined by radioimmunoassay [9].…”

Extracorporeal perfusion of isolated organs of large animals – Bridging the gap between in vitro and in vivo studies

“…Systems which employ a high flow approach aim for physiological flow rates and perfusion pressures to mimic the in vivo situation more closely. As a higher flow rate leads to lower organ resistance, this indicates the integrity of blood vessels (Barthel et al, 1989;Bristol et al, 1991;Wagner et al, 2003). Lactate levels in venous perfusate samples obtained during high flow experiments are lower than in low flow experiments and this could also imply a better oxygenation of tissues (Wagner et al, 2003).…”

“…Reported diluents have been cell free solutions (Domingo-Pech et al, 1991;Wagner et al, 2003;Erasmus et al, 2006;Friebe et al, 2013b) or autologous plasma (Patan et al, 2009;Patan-Zugaj et al, 2012, which comes with the added benefit of increasing oncotic pressure. As cell free solutions are associated with low oncotic pressure, resulting in edema formation and weight gain, adding plasma expanders such as plasma proteins (Verbeke et al, 1968;Roets et al, 1974;Patan et al, 2009) and purified albumin (Rehfeld et al, 1982;Barthel et al, 1989;Riviere et al, 1989;Brunicardi et al, 2001) perfused specimens thus reflects the degree of ischemia/reperfusion injury (Petrasek et al, 1994;Adham et al, 1997) and the integrity of the microvasculature (Müller et al, 2013). Excessive edema formation results in rapidly progressive deterioration of organ function, which renders it unsuitable for transplantation and research alike (Verbeke et al, 1972).…”