A novel paracorporeal method for isolated rodent lung reperfusion.
BACKGROUND
The isolated perfused lung model is commonly used in small animals to study lung function after preservation and cold storage. Measurements of oxygenation, compliance, and capillary filtration coefficient (Kf) permit analysis of preservation solutions or modifications of these solutions. However, inter-investigator variability using different perfusates makes comparisons difficult. Whole blood perfusion more closely mimics the in vivo situation, but extracorporeal circulation may alter the physiologic integrity of the model. Paracorporeal support has been used, but this technique required mechanical ventilation of the support rodent and did not incorporate a method for determining Kf. We evaluated a less-invasive technique, of providing cross-circulatory syngeneic support, maintaining the ability to compute Kf.


METHODS
Angiocatheters were inserted into both femoral arteries and one femoral vein of the support rat. The venous cannula was connected to the pulmonary artery of the ex vivo lung block to provide inflow. Pulmonary effluent blood from the lung block was collected via a left atrial cannula and returned to the support rat via the femoral artery. A separate, height-adjustable column was included in the circuit for measurement of Kf.


RESULTS
Each support rat was used to sequentially perfuse three double-lung blocks. The inflow sample to each lung block was analyzed for pH, pO2, pCO2, and hematocrit to follow alterations in support rat physiology. There were no statistical differences in the pH, PO2, or hematocrit. No significant differences were noted in the pO2 of the pulmonary effluent blood or the Kf; analyzed to determine whether the sequence of reperfusion affected the pulmonary function assessment.


CONCLUSIONS
The syngeneic support rat delivers constant pressure systemic venous blood at stable physiologic parameters to the ex vivo lung block. Recirculation of the perfusate through the support rat diminishes the need to pool blood from donors, detoxifies and deoxygenates pulmonary effluent blood, and permits examination of sequential lung blocks. This technique represents a hybrid model between isolated perfused and orthotopic transplant models, maintaining Kf determination, a sensitive indicator of reperfusion injury. This technique could be applicable to reperfusion injury models of other organs (using arterial inflow instead) and may permit increased standardization among investigators.
Baker
C J
CJ
Department of Cardiothoracic Surgery, University of Southern California and Childrens Hospital, Los Angeles 90033, USA.
Quardt
S M
SM
Kim
J D
JD
Darbinian
S H
SH
Starnes
V A
VA
Barr
M L
ML
D000818:Animals; D001784:Blood Gas Analysis; D005112:Extracorporeal Circulation / Q000379:methods*; D008168:Lung / Q000098:blood supply* / Q000502:physiology; D008297:Male; D023421:Models, Animal; D051381:Rats; D011917:Rats, Inbred Lew; D015424:Reperfusion / Q000379:methods; D014021:Tissue Preservation