Infusing Mesenchymal Stromal Cells into Porcine Kidneys during Normothermic Machine Perfusion: Intact MSCs Can Be Traced and Localised to Glomeruli.

Merel Pool,Marco Eijken,Henri Leuvenink,Bente Jespersen,Marlies Reinders,Cyril Moers,Martin Hoogduijn,Rutger Ploeg,Nils ’T Hart,Jesus Sierra Parraga,Tim Eertman,Marieke Roemeling-Van Rhijn

doi:10.3390/ijms20143607

Abstract

Normothermic machine perfusion (NMP) of kidneys offers the opportunity to perform active interventions, such as the addition of mesenchymal stromal cells (MSCs), to an isolated organ prior to transplantation. The purpose of this study was to determine whether administering MSCs to kidneys during NMP is feasible, what the effect of NMP is on MSCs and whether intact MSCs are retained in the kidney and to which structures they home. Viable porcine kidneys were obtained from a slaughterhouse. Kidneys were machine perfused during 7 h at 37 °C. After 1 h of perfusion either 0, 105, 106 or 107 human adipose tissue derived MSCs were added. Additional ex vivo perfusions were conducted with fluorescent pre-labelled bone-marrow derived MSCs to assess localisation and survival of MSCs during NMP. After NMP, intact MSCs were detected by immunohistochemistry in the lumen of glomerular capillaries, but only in the 107 MSC group. The experiments with fluorescent pre-labelled MSCs showed that only a minority of glomeruli were positive for infused MSCs and most of these glomeruli contained multiple MSCs. Flow cytometry showed that the number of infused MSCs in the perfusion circuit steeply declined during NMP to approximately 10%. In conclusion, the number of circulating MSCs in the perfusate decreases rapidly in time and after NMP only a small portion of the MSCs are intact and these appear to be clustered in a minority of glomeruli.

Highlights

In an attempt to decrease waiting time for a donor kidney, donation after circulatory death (DCD) is starting to play a more prominent role in many transplant centres [1]
The purpose of this study was to determine whether administering mesenchymal stromal cells (MSCs) during Normothermic machine perfusion (NMP) is technically feasible, what the effect of NMP is on the survival of MSCs, whether intact MSCs can be detected in the kidney after NMP and to which renal structures these cells localise
The dark areas in the renal cortex represent the MSCs. This is the first study in which human MSCs were administered during NMP of porcine kidneys

Summary

Introduction

In an attempt to decrease waiting time for a donor kidney, donation after circulatory death (DCD) is starting to play a more prominent role in many transplant centres [1]. Preserving the function of ischaemically damaged kidney grafts is of vital importance for an effective transplantation. Pre-transplant machine perfusion could enable active organ conditioning and provides a platform for interventions prior to transplantation [4]. This is preferably done under near-physiological conditions, through (sub)normothermic machine perfusion at or just below 37 ◦C [5]. Normothermic machine perfusion (NMP) restores organ metabolism outside the body in absence of an allogeneic immune response prior to transplantation and this may reverse some effects of ischaemia. NMP could provide a platform for active interventions to an isolated organ prior to transplantation, such as administering cellular therapy

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Conclusion