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

• Published in: International journal of molecular sciences Vol. 20; no. 14; p. 3607
• Main Authors: Pool, Merel, Eertman, Tim, Sierra Parraga, Jesus, 't Hart, Nils, Roemeling-van Rhijn, Marieke, Eijken, Marco, Jespersen, Bente, Reinders, Marlies, Hoogduijn, Martin, Ploeg, Rutger, Leuvenink, Henri, Moers, Cyril
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 23.07.2019; MDPI
• Subjects: Adipocytes - cytology; Adipocytes - physiology; Adipose tissue; Animals; Biopsy; Bone marrow; Bone Marrow Cells - cytology; Bone Marrow Cells - physiology; CD105 antigen; CD45 antigen; CD73 antigen; CD90 antigen; Cell Differentiation; Cell Tracking - methods; Endothelial cells; Experiments; Flow cytometry; Fluorescence; Fluorescent Dyes - metabolism; Hemodynamics; Humans; Kidney Glomerulus - surgery; Kidney Glomerulus - ultrastructure; Kidney Transplantation; Kidney transplants; Kidneys; Lymphocytes; Markers; Mesenchymal Stem Cell Transplantation - instrumentation; Mesenchymal Stem Cell Transplantation - methods; Mesenchymal Stem Cells - cytology; Mesenchymal Stem Cells - physiology; mesenchymal stromal cells; Mesenchyme; Microscopy; Microscopy, Fluorescence; normothermic machine perfusion; organ preservation; Organ Preservation - methods; Organic Chemicals - metabolism; Perfusion; Perfusion - instrumentation; Perfusion - methods; porcine; Stains & staining; Stromal cells; Swine; Temperature; Transplantation; Transplantation, Heterologous; Transplants & implants; Urine; porcine; normothermic machine perfusion; kidney transplantation; mesenchymal stromal cells; organ preservation
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms20143607

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, 10 , 10 or 10 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 10 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.