Effective delivery of Mesenchymal Stromal cells during isolated liver machine perfusion to promote graft repair on the pump

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M.M.A. Verstegen, W. Wang, L. Mezzanotte, J.P. van Kooten, S. van den Hoek, I. Schurink, P. de Ruiter, R. Yanto Ridwan, M.J. Hoogduijn, J.M. Sierra Parraga, C.W.G.M. Löwik, J.E. de Haan, P.A.C. de Specht, L.J.W. van der Laan, J. de Jonge

Chair(s): Dr. B.G. Hepkema

Thursday 9 march 2017

12:30 - 12:45h

Categories: Poster - Basaal

Parallel session: Postersessie - Basaal 1

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Faced with current organ shortages more donor with compromised graft quality are being used. These grafts have less favorable outcome after transplantation due to increased graft damage. Currently machine preservation provides new opportunities, not only to improve graft preservation, but moreover to apply regenerative medicine strategies to repair damaged organs. Mesenchymal Stromal cells (MSCs) represent a potential new therapeutic strategy to stimulate liver regeneration. The aim of the current study is to investigate whether MSCs can be effectively delivered during hypothermic oxygenated machine perfusion of porcine liver grafts.

Livers from Yorkshire mini pigs (n=9, age 2-3 months, female, 20-30 kg) were procured according to standard methods and cannulated, cooled and flushed with cold preservation before infusion with human bone marrow-derived MSCs. MSCs (10 to 15 million per infusion) were genetically labeled with a luciferase reporter gene to visualize the distribution of cells after infused using bioluminescence imaging.  The livers were perfused for 1h at 10˚C with oxygenated preservation fluid before warm, oxygenated reperfusion was done with the pig’s own blood. The infused MSCs were identified in biopsies using human specific q-PCR and immunohistochemistry  for CK19. Cytokine production of the infused MSC after 4h of warm reperfusion was measured using the Luminex Multiplex platform.

MSCs were clearly visualized directly after infusion (t=0) and following  30 min of cold, oxygenated perfusion. Whole organ imaging showed an even distribution of cells throughout the liver graft, when infused in the hepatic artery and an even spread throughout the liver after 30 min of perfusion. The delivery of human MSC  to the graft was confirmed by human specific q-PCR of multiple biopsy samples and by CK19 staining. After 4 h of warm reperfusion with the pigs blood, human-specific cytokines IL-6 and IL-8 were detected in the pig serum, demonstrating that the infused MSC were functionally active during the warm reperfusion.

Conclusion:
human MSCs can be effectively delivered to ischemic liver grafts during hypothermic oxygenated machine perfusion. This study shows feasibility of MSC-based therapies for repair of damaged organs on the pump, which is currently investigated with human livers discarded for transplantation.