Evaluation of a system for the perfusion of isolated, rodent organs

Perfusion of isolated organs is a common experimental approach. However, the surfaces of the perfusion system might alter the components of the blood and thereby negatively affect organ function. The aim of this study was to minimize the influence of the perfusion system on the blood components and to evaluate the system. Pressure and flow in the perfusion system consisting of a roller-pump, reservoir, oxygenator, hemo-filter and bubble-trap with a total tubing length of 4.5 m are controlled by a computer software (DASYLAB, Datalog, Moenchengladbach, Germany) via a transducer connected to the system. The organ to be perfused is positioned under a microscope (Orthoplan, Leica, Bensheim, Germany), allowing the investigation of microcirculatory parameters. The images raised are recorded on video tapes. To evaluate the system it was perfused with human blood (Hct 28 to 30%) for 90 min. Heparin (n = 6) or citrate (n = 6) served as anti-coagulants. The disappearance of cells from the blood was determined at time points 0, 1, 5, 10, 15, 20, 30, 45, 60, 75 and 90 min by means of a cell counter (AC T8, Coulter Beckmann, Krefeld, Germany). Cell activation was assessed by analysis of the expression of L- and P-selectin and CD11b. The activation of the complement system was examined by measuring the serum levels of the complement factors C3c and C4. There was no significant loss or activation of the blood cells at any of the above given time points. The serum levels of the complement factors remained within the physiological range and showed no changes throughout the whole experiments. Thus, the perfusion system does not have a negative influence on the blood and its individual components, and is therefore a reliable tool for perfusion experiments.