Methods in Bioengineering: Organ Preservation and Reengineering -- Contents -- Preface -- 1 Normothermic Machine Perfusion of the Liver -- 1.1 Introduction -- 1.1.1 Organ Preservation -- 1.1.2 Cold Storage Preservation -- 1.1.3 Normothermic Machine Perfusion -- 1.1.4 Viability Testing -- 1.1.5 Experimental Data -- 1.1.6 Other Fields of Application -- 1.2 Experimental Design and Methods -- 1.2.1 Materials -- 1.2.2 Methods -- 1.2.3 Parameters -- 1.3 Data Acquisition-Anticipated Results and Interpretation -- 1.3.1 Oxygenation/Blood Gas Analysis -- 1.3.2 Pressure Profile -- 1.3.3 Laboratory Values -- 1.3.4 Histology -- 1.3.5 Immunohistochemistry -- 1.3.6 Transplantation -- 1.4 Discussion and Commentary -- References -- 2 Heart Preservation: Conventional Versus Novel Techniques -- 2.1 Introduction -- 2.1.1 Cold Storage -- 2.1.2 Continuous Perfusion of the Donor Heart -- 2.1.3 Hypothermic Continuous Perfusion -- 2.1.4 Potential Advantages of CP -- 2.1.5 Genetic Therapy: Targeting the Donor Heart -- 2.1.6 Disadvantages of CP -- 2.2 Materials and Methods -- 2.2.1 Materials -- 2.2.2 Methods -- 2.2.2 Graft Assessment -- 2.3 Discussion and Commentary -- 2.4 Summary Points -- References -- 3 Hypothermic Machine Perfusion of Kidneys -- 3.1 Introduction -- 3.1.1 Basic Principles of Organ Preservation -- 3.1.2 History of Hypothermic Perfusion -- 3.1.3 Need for Machine Perfusion in Transplantation -- 3.2 Experimental Methods -- 3.2.1 Hypothermic Perfusion Preservation (4-7°C) -- 3.2.2 Midthermia Perfusion Preservation (15-20°C) -- 3.3 Discussion and Commentary -- 3.4 Application Notes -- 3.5 Summary Points -- References -- 4 Hypothermic Machine Perfusion of Livers -- 4.1 Introduction -- 4.2 Experimental Design -- 4.3 Materials -- 4.3.1 Rat Experiments -- 4.3.2 Swine Experiments -- 4.4 Methods -- 4.4.1 Rat Experiments -- 4.4.2 Swine Experiments.

4.5 Data Analysis -- 4.6 Anticipated Results -- 4.7 Discussions and Commentary -- 4.8 Application Notes -- 4.9 Summary Points -- Acknowledgments -- References -- 5 Hypothermic Perfusion of Pancreas: Emphasis on Preservation Prior to Islet Isolation -- 5.1 Introduction -- 5.2 Experimental Methods and Materials -- 5.2.1 Materials -- 5.2.2 Methods -- 5.3 Data Acquisition and Anticipated Results -- 5.4 Discussion and Commentary -- 5.4.1 Pancreas Perfusion on the LifePort Kidney Transporter -- 5.4.2 Pancreas Cannulation for Perfusion -- 5.4.3 Adaptation for Future Applications -- 5.5 Troubleshooting -- 5.6 Application Notes -- 5.7 Summary Points -- Acknowledgments -- References -- 6 Methods of Cardiac Oxygen Persufflation -- 6.1 Introduction -- 6.1.1 Importance of Oxygenation and Other Technical Features in Organ Preservation -- 6.1.2 Development of Oxygenation in Organ Preservation Techniques Without Continuous Fluid Perfusion -- 6.1.3 Heart Preservation-Optimal Oxygenation Without Continuous Fluid Perfusion -- 6.2 Materials -- 6.2.1 Production of the Valve Guard -- 6.3 Methods -- 6.4 Data Acquisition and Results -- 6.4.1 14-Hour Hypothermic Heart Storage Experiments -- 6.4.2 NHBD Heart Experiments with 16-Minute Normothermic Ischemia and 3.3 Hours of Hypothermic Preservation -- 6.4.3 Evaluation of Coronary Endothelial Function After Preservation and Transplantation of NHBD Hearts -- 6.4.4 Evaluation of Coronary Resistance Vessel After Prolonged COP -- 6.4.5 Evaluation of Metabolic Effect of COP During 3-Hour Preservation After 15-Minute NHBD Phase -- 6.4.6 7-Day Recovery After 14-Hour Persufflation and Heterotopic Transplantation -- 6.5 Discussion and Commentary -- 6.6 Summary Points -- Acknowledgments -- References -- 7 Vascular Oxygen Persufflation for Preservation and Reconditioning of Marginal Liver Grafts -- 7.1 Introduction -- 7.2 Materials.

7.3 Methods -- 7.3.1 Surgery -- 7.3.2 Device Setup -- 7.3.3 Connecting Liver/Starting Persufflation -- 7.3.4 Graft Evaluation -- 7.4 Discussion and Commentary -- 7.5 Application Notes -- 7.5.1 Donation After Cardiac Death (DCD) -- 7.5.2 Fatty Livers -- 7.5.3 Hypothermic Reconditioning (HR) for Conventionally Stored Organs -- 7.6 Summary Points -- References -- 8 Use of Optical Imaging and Spectroscopy in Assessment of Organ Perfusion -- 8.1 Introduction -- 8.1.1 Spectral Imaging Technologies Overview -- 8.2 Experimental Design -- 8.3 Materials -- 8.4 Methods -- 8.4.1 Room Setup -- 8.4.2 Preparation for Kidney Harvest -- 8.4.3 Left Kidney Harvest, Preservation, and Data Collection -- 8.4.4 Autotransplantation -- 8.4.5 Reanastomosis -- 8.4.6 Completion of Surgery, Recovery, and Euthanasia -- 8.5 Data Acquisition, Anticipated Results, and Interpretation -- 8.5.1 3-CCD -- 8.5.2 Infrared -- 8.5.3 VRIS -- 8.6 Discussion and Commentary -- 8.7 Summary Points -- Acknowledgments -- Disclaimer -- References -- 9 Preparation of a Transplantable Liver Graft by the Recellularization of a Decellularized Whole Organ -- 9.1 Introduction -- 9.2 Methods -- 9.2.1 Liver Harvest -- 9.2.2 Perfusion Decellularization -- 9.2.3 Recellularization and In Vitro Perfusion Culture of the Recellularized Graft -- 9.2.4 Heterotopic Transplantation -- 9.3 Anticipated Results -- 9.3.1 Decellularization -- 9.3.2 Recellularization -- 9.3.3 Heterotopic Transplantation -- 9.4 Discussion and Commentary -- Acknowledgments -- References -- 10 Engineering Lung Tissue -- 10.1 Introduction -- 10.2 Experimental Design -- 10.3 Materials -- 10.3.1 Reagents -- 10.3.2 Facilities and Equipment -- 10.4 Methods -- 10.4.1 Bioreactor Assembly -- 10.4.2 Organ Harvest -- 10.4.3 Organ Decellularization -- 10.4.4 Organ Rinsing and Sterilization -- 10.4.5 Recellularization -- 10.4.6 Organ Culture.

10.4.7 Staining, Blotting, and Data Acquisition -- 10.5 Anticipated Results -- 10.6 Discussion and Commentary -- 10.7 Application Notes -- 10.8 Summary Points -- Acknowledgments -- References -- 11 Detergent-Enzymatic Method for Bioengineering Human Airways -- 11.1 Introduction -- 11.2 Experimental Design -- 11.3 Reagents -- 11.3.1 Preparation of a Human Airway Matrix: Detergent-Enzymatic Protocol -- 11.3.2 Preparation of a Human Airway Matrix: Histology, Immunohistology, and Immunofluorescence Histology -- 11.4 Methods -- 11.4.1 Preparation of a Human Airway Matrix -- 11.4.2 Preparation and Differentiation of Chondrocyte Cultures -- 11.4.3 Preparation of Epithelial Cultures -- 11.4.4 Characterization Study -- 11.4.5 Bioreactor Design -- 11.4.6 Graft Preparation -- 11.5 Anticipated Results -- 11.6 Discussion and Commentary -- 11.7 Clinical Notes -- 11.8 Summary Points -- References -- 12 Ex Vivo Perfusion of Rat Liver Grafts for Hepatocyte Retrieval -- 12.1 Introduction -- 12.2 Experimental Design -- 12.2.1 Systems Requirements -- 12.3 Methods -- 12.3.1 Blood Harvest -- 12.3.2 Liver Harvest -- 12.3.3 Warm Ischemia and Cuffing -- 12.3.4 Liver Perfusion -- 12.3.5 Hepatocyte Isolation of Fresh and Perfused Livers -- 12.4 Data Acquisition -- 12.5 Anticipated Results -- 12.6 Discussion and Commentary -- 12.7 Application Notes -- 12.8 Summary Points -- Acknowledgments -- References -- 13 Technique of Clinical Vascular Isolation and Perfusion of the Liver -- 13.2 Typical Patient Experience -- 13.3 Materials -- 13.4 Methods -- 13.4.1 Surgical Preparation of the Liver for Isolation -- 13.4.2 Establishment of the Isolated Hepatic Perfusion Circuit -- 13.4.3 Operation of the Isolated Perfusion Circuit and Leak Monitoring -- 13.4.4 Establishment of the Normal Liver Perfusion and Termination of the Procedure.

13.5 Data Acquisition, Interpretation, and Patient Management During Perfusion -- 13.6 Anticipated Results -- 13.7 Discussion and Commentary -- 13.8 Application Notes -- 13.9 Summary Points -- References -- 13.1.1 The Unique Aspects of Arterial and Venous Liver Anatomy -- 13.1.2 Rationale for the Use of IHP in Clinical Oncology -- 13.1.3 History and Initial Clinical Results with IHP -- About the Editors -- Index.