Treating ischemically damaged porcine kidneys with mesenchymal stromal cells during normothermic machine perfusion

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M.B.F. Pool, J.M. Sierra Parraga, M. Roemeling-van Rhijn, M.E.J. Reinders, M.J. Hoogduijn, R.J. Ploeg, H.G.D. Leuvenink, C. Moers

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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Donor kidneys of inferior quality are increasingly being accepted to decrease waiting time for a transplant. Normothermic machine perfusion (NMP) could provide superior organ preservation, compared to cold preservation methods and may also be a tool for pre-transplant quality assessment of such marginal grafts. In addition, NMP offers the unique opportunity for active interventions to an isolated organ under near-physiological conditions prior to transplantation. There is increasing evidence that mesenchymal stromal cells (MSCs) could have a positive effect on ischaemia-reperfusion (IR) injury. However, in most studies MSCs are administered to recipients after transplantation, which exposes the whole patient to circulating allogeneic cells. Moreover, administering MSCs earlier in the IR cascade might create better opportunities for beneficial effects. The purpose of this study was to determine whether administering MSCs during NMP is technically feasible, if MSCs reach the kidney and remain viable, to which structures they home and which cytokines are secreted by MSCs during NMP.

Porcine kidneys and autologous blood were obtained from two slaughterhouses. Warm ischaemia time was standardised at 30 min and cold ischaemia time was 3.5-5 hours. Kidneys were machine perfused in a recirculating circuit with 350 ml washed autologous red blood cells, 500 ml Williams’ Medium E, albumin, creatinine and Augmentin during 6 hours at 37°C. After 1 hour of perfusion either 0, 10^5, 10^6 or 10^7 cultured human adipose tissue derived MSCs were added (n=3 per group). Vital parameters were monitored and perfusate and urine samples were taken regularly. Biopsies were taken to assess renal histology and to locate MSCs with immunohistochemistry. Luminex analysis was used to determine secretion products of MSCs.

After NMP vital MSCs were detected in the lumen of glomerular capillaries in the 10^7 MSC group, but not in the other groups. MSCs secreted pro-inflammatory cytokines IL-6, IL-8 and MCP-1 in response to the ischemically damaged kidney during NMP, in a dose-dependent fashion.

In conclusion, adding MSCs during pre-transplant renal NMP proved to be feasible. MSCs remained viable and detectable and secreted various pro-inflammatory cytokines. It remains to be studied if this leads to beneficial or harmful processes for the organ, which molecular pathways are activated by exposure to MSCs and whether MSC pre-treatment will indeed enhance renal function post-transplant.