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University Health Network, Toronto General Hospital

11 ARTICLES PUBLISHED IN JoVE

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Medicine

Technique of Subnormothermic Ex Vivo Liver Perfusion for the Storage, Assessment, and Repair of Marginal Liver Grafts
Jan M. Knaak *1,2, Vinzent N. Spetzler *1,2, Nicolas Goldaracena 1,2, Kristine S. Louis 1,2, Nazia Selzner 1,3, Markus Selzner 1,2
1Multi Organ Transplant Program, Toronto General Hospital, 2Department of Surgery, Toronto General Hospital, 3Department of Medicine, Toronto General Hospital

Marginal grafts, such as fatty livers, grafts from older donors, or livers retrieved after cardiac death (DCD) tolerate conventional, cold static storage only poorly. We developed a novel model of subnormothermic ex vivo liver perfusion for preservation, assessment, and repair of marginal liver grafts prior to transplantation.

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Medicine

Technique of Porcine Liver Procurement and Orthotopic Transplantation using an Active Porto-Caval Shunt
Vinzent N. Spetzler 1, Nicolas Goldaracena 1, Jan M. Knaak 1, Kristine S. Louis 1, Nazia Selzner 1, Markus Selzner 1
1Multi Organ Transplant Program, Department of Surgery, Toronto General Hospital

Experimental animal research plays a pivotal role in the development of clinical transplantation practice. The porcine orthotopic liver transplantation model (OLTx) closely resembles human conditions and is frequently used in clinically oriented research. The following protocol contains all information for a reliable porcine OLTx model using an active porto-caval-jugular shunt.

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Medicine

Normothermic Ex Vivo Kidney Perfusion for the Preservation of Kidney Grafts prior to Transplantation
J. Moritz Kaths 1,2, Vinzent N. Spetzler 1, Nicolas Goldaracena 1, Juan Echeverri 1, Kristine S. Louis 1, Daniel B. Foltys 3, Mari Strempel 4, Paul Yip 5, Rohan John 5, Istvan Mucsi 1, Anand Ghanekar 1, Darius Bagli 6,7, Lisa Robinson 2, Markus Selzner 1
1Multi Organ Transplant Program, Department of Surgery, Toronto General Hospital, 2Division of Nephrology, The Hospital for Sick Children, Toronto, 3Department of General, Visceral & Transplant Surgery, University Medical Center Mainz, 4Department of Abdominal, Vascular & Transplant Surgery, Merheim Medical Center Cologne, 5Laboratory Medicine & Pathobiology, Toronto General Hospital, 6Departments of Surgery (Urology) & Physiology, The Hospital for Sick Children, Toronto, 7Developmental & Stem Cell Biology, The Hospital for Sick Children, Toronto

The severe organ shortage has resulted in increased use of marginal kidney grafts for transplantation. This has triggered interest in alternative storage methods, since marginal grafts especially tolerate cold storage poorly. The technique of normothermic ex vivo kidney perfusion (NEVKP) represents a novel preservation method for kidney grafts prior to  transplantation.

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Medicine

Heterotopic Renal Autotransplantation in a Porcine Model: A Step-by-Step Protocol
J. Moritz Kaths 1,2, Juan Echeverri 1,3, Nicolas Goldaracena 1, Kristine S. Louis 1, Paul Yip 4, Rohan John 4, Istvan Mucsi 5, Anand Ghanekar 1, Darius Bagli 6, Markus Selzner 1, Lisa A. Robinson 2
1Multi Organ Transplant Program, Department of Surgery, Toronto General Hospital, 2Division of Nephrology, The Hospital for Sick Children, 3Programa de Doctorat en Medicina, La Universitat Autónoma de Barcelona, 4Laboratory Medicine and Pathobiology, Toronto General Hospital, 5Department of Medicine, Toronto General Hospital, 6Departments of Surgery (Urology) & Physiology, Developmental & Stem Cell Biology, The Hospital for Sick Children

Porcine models of organ transplantation provide an important platform to study mechanisms of organ preservation. This article describes a heterotopic porcine renal autotransplantation model, which allows investigating new approaches to improve the outcome of transplantation using marginal kidney grafts.

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Neuroscience

Microstate and Omega Complexity Analyses of the Resting-state Electroencephalography
Fei Gao 1, Huibin Jia 2, Yi Feng 1
1Department of Pain Medicine, Peking University People's Hospital, 2Key Laboratory of Child Development and Learning Science of Ministry of Education, Research Center for Learning Science, School of Biological Sciences & Medical Engineering, Southeast University

This article describes the protocol underlying electroencephalography (EEG) microstate analysis and omega complexity analysis, which are two reference-free EEG measures and highly valuable to explore the neural mechanisms of brain disorders.

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Medicine

Using a Chemical Biopsy for Graft Quality Assessment
Iga Stryjak *1, Natalia Warmuzińska *1, Kamil Łuczykowski 1, Matyas Hamar 2, Peter Urbanellis 2, Emilia Wojtal 3, Marek Masztalerz 3, Markus Selzner 2,4, Zbigniew Włodarczyk 3, Barbara Bojko 1
1Department of Pharmacodynamics and Molecular Pharmacology, Faculty of Pharmacy, Nicolaus Copernicus University in Torun, Collegium Medicum in Bydgoszcz, 2Multi Organ Transplant Program, Department of Surgery, Toronto General Hospital, University Health Network, 3Department of Transplantology and General Surgery, Collegium Medicum in Bydgoszcz, Antoni Jurasz University Hospital No. 1 in Bydgoszcz, Nicolaus Copernicus University in Torun, Bydgoszcz, Poland, 4Department of Medicine, Toronto General Hospital

The protocol presents utilization of the chemical biopsy approach followed by comprehensive metabolomic and lipidomic analysis for quality assessment of kidney grafts allocated for transplantation.

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Medicine

Normothermic Ex Vivo Pancreas Perfusion for the Preservation of Pancreas Allografts before Transplantation
Catherine Parmentier 1, Samrat Ray 1, Laura Mazilescu 1,2, Masataka Kawamura 1, Yuki Noguchi 1, Emmanuel Nogueira 1, Sujani Ganesh 1, Bhranavi Arulratnam 1, Sangeetha Kalimuthu 3, Markus Selzner 1, Trevor Reichman 1
1Ajmera Transplant Program, University Health Network, Toronto General Hospital, 2Department of General, Visceral, and Transplantation Surgery, University Hospital Essen, 3Department of Pathology, University Health Network, Toronto General Hospital

Normothermic ex vivo machine perfusion (NEVP) has scarcely been explored for the preservation of pancreas allografts. We present an innovative preservation technique for pancreas allografts before transplantation.

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Bioengineering

Procurement and Perfusion-Decellularization of Porcine Vascularized Flaps in a Customized Perfusion Bioreactor
Michael S. Xu 1, Golnaz Karoubi 1,2, Thomas K. Waddell 1,3, Siba Haykal 1,4
1Latner Thoracic Surgery Research Laboratories, Toronto General Hospital Research Institute, University Health Network, 2Institute of Laboratory Medicine and Pathobiology, University of Toronto, 3Division of Thoracic Surgery, Department of Surgery, University of Toronto, 4Division of Plastic and Reconstructive Surgery, Department of Surgery, University of Toronto

The protocol describes the surgical procurement and subsequent decellularization of vascularized porcine flaps by the perfusion of sodium dodecyl sulfate detergent through the flap vasculature in a customized perfusion bioreactor.

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Bioengineering

Procurement and Decellularization of Rat Hindlimbs Using an Ex Vivo Perfusion-Based Bioreactor for Vascularized Composite Allotransplantation
Aisha Adil 1,2, Golnaz Karoubi 1,3, Siba Haykal 1,2,4
1Latner Thoracic Research Surgical Laboratories, University Health Network, Toronto General Hospital, 2Institute of Medical Science, Temerty Faculty of Medicine, University of Toronto, 3Institute of Laboratory Medicine and Pathobiology, Temerty Faculty of Medicine, University of Toronto, 4Division of Plastic & Reconstructive Surgery, Department of Surgery, University of Toronto

We describe the surgical technique and decellularization process for composite rat hindlimbs. Decellularization is conducted using low-concentration sodium dodecyl sulfate through an ex vivo machine perfusion system.

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Medicine

Surgical Tips and Tricks for Performing Porcine Pancreas Transplantation
Samrat Ray 1, Catherine Parmentier 1, Laura Mazilescu 1,2, Masataka Kawamura 1, Yuki Noguchi 1, Emmanuel Nogueira 1, Sujani Ganesh 1, Bhranavi Arulratnam 1, Markus Selzner 1, Trevor Reichman 1
1Ajmera Transplant Centre, University Health Network, Toronto general hospital, 2Department of general, visceral, and transplantation surgery, University Hospital Essen

The video article summarizes the technique of pancreatectomy and pancreas allotransplantation in a porcine 3-day survival model with a step-by-step description of the method and emphasis on the surgical tips and tricks to deal with the precarious and delicate porcine visceral anatomy.

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Medicine

Intubation, Central Venous Catheter, and Arterial Line Placement in Swine for Translational Research in Abdominal Transplantation Surgery
Catherine Parmentier 1, Fei Gao 1, Samrat Ray 1, Masataka Kawamura 1, Emmanuel Noguiera 1, Sujani Ganesh 1, Markus Selzner 1, Trevor Reichman 1
1Ajmera Transplant Program, University Health Network, Toronto General Hospital

To obtain the best possible results, surgeons performing translational research experiments should be proficient at intubation and vascular line placement. This paper describes the techniques used by the Toronto Organ Preservation Laboratory to perform these procedures.

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