Abstract
Single lung transplantation is an acceptable therapeutic option. Here we use our single lung porcine allograft as a vehicle for stem cell transfer. We present our protocol for perfusion of porcine lungs with human stem cells using our ex-vivo machine.
Adult pigs underwent unilateral left pneumonectomy. The explanted lungs were removed from surgical field and connected to ex-vivo circuit. Porcine lungs were perfused with human albumin after explanation. Lungs were ventilated during perfusion. Total ex vivo perfusion <3 hours. 2 methods of BMC administration were compared:
In group 1 CD133+ cells were only passed through the needle. In group2, MSCs were perfused into the pulmonary artery using the ex vivo machine
1. Human male somatic CD133+ Cells were passed through a Microlance needle (BD Gr.20, ∅ 0,8 × 40 mm, 26 Gauge)without injection into the lung. 750,000 human CD133+ cells (mobilized peripheral blood), purity (92.4-99.4%), frozen sample were used and different verification runs pump/syringe (n=10, flow rate up to 4mL/min, 30sec), viability was not unchanged (Table 1). 2. 100,000,000 human BMCs were perfused using the ex-vivo machine. The viability of the cells before and after lung perfusion did not show any significant in regard to viability rate, FACS and histological characteristics.
Both methods showed safe application route for pulmonary stem cell transfer. The viability of CD105+/45- cells is not as affected during ex vivo perfusion as rest of the cells. The Ex-Vivo mediated transfer is more practical for lung treatment. More histologic and cell culture studies are needed to confirm the feasibility.
