Pancreatic Islet Isolation: From the Mouse to the Clinic (Advances in Experimental Medicine and Biology Book 938)

Pancreatic Islets Make Up The Endocrine Pancreas And They Contain The Only Source Of Insulin In The Body, Beta Cells. Hence, Access To High Quality Preparations Of Pancreatic Islets Is Fundamental For In Vitro Studies And To Test Pre-clinical Applications In Animal Models In Vivo. Access To Healthy Human Islets Is Also Crucial To Improve Transplantation Procedures For Diabetes. Given The Susceptibility Of Pancreatic Islets To The Enzymatic Digestion And Mechanical Stress Required To Obtain Them, The Isolation Of Islets Is Often Considered As The Delicate Work Of A Craftsman. This Book, Which Is Aimed At Beginners And Experts Alike, Is A Survey Of The Current State-of?the-art In This Field And It Centres On The Challenges, Pitfalls And Peculiarities Of Pancreatic Islet Isolation In The Different Species Used In Pre-clinical And Clinical Applications. It Explores The Similarities And Differences Between Human Islets And Those From Other Relevant Species (rodents, Pigs And Non-human Primates), And How These Influence Islet Isolation. The Ultimate Goal Of This Book Is To Improve The Outcome Of Islet Isolation And Transplantation In Pre-clinical And Clinical Applications. Dedication; Preface; Contents; Contributors; 1: Historical Background Of€pancreatic Islet Isolation; 1.1 The History Of€diabetes Mellitus; 1.1.1 Discovery Of€insulin; 1.2 The History Of€islet Isolation; 1.3 Concluding Remarks; References; 2: The Different Faces Of€the€pancreatic Islet; 2.1 Introduction; 2.1.1 Cytoarchitecture; 2.1.2 Vasculature; 2.1.3 Paracrine Signaling; 2.1.4 Innervation; 2.2 Summary; References; 3: Isolation Of€mouse Pancreatic Islets Of€langerhans; 3.1 Introduction; 3.2 Pancreas Distention And€dissection; 3.2.1 General Considerations About€the€procedure. 3.2.2 Enzyme Administration3.3 Pancreas Digestion; 3.3.1 General Considerations About€the€procedure; 3.3.2 Selection Of€the€enzyme; 3.3.3 Duration Of€digestion; 3.4 Islet Purification; 3.4.1 General Considerations About€the€procedure; 3.4.2 Density Gradient Purification; 3.4.3 Other Purification Methods; 3.5 Islet Culture; 3.5.1 General Considerations About€the€procedure; 3.6 Concluding Remarks; References; 4: Pancreatic Islets: Methods For€isolation And€purification Of€juvenile And€adult Pig Islets; 4.1 Introduction; 4.2 Pig Donor Variables; 4.2.1 Pig Strain. 4.2.2 Porcine Donor Age4.3 Pig Pancreas Procurement; 4.3.1 Intraductal Pancreas Perfusion Technique; 4.3.2 Intraductal Pancreas Perfusion Buffer; 4.4 Pig Pancreas Digestion; 4.4.1 Enzyme Blend Selection; 4.4.2 Dissociation; 4.5 Porcine Islet Purification; 4.5.1 Determinants Of€intrinsic Pig Pancreas Density; 4.5.2 Reversing Cell Swelling; 4.5.3 Selection Of€density Gradient Media; 4.5.4 Density Gradient Centrifugation Technique; 4.5.5 Isokinetic Islet Purification; 4.6 Porcine Islet Culture; 4.6.1 Selecting The€most Suitable Culture Medium; 4.6.2 Glucose Concentration. 4.6.3 Serum Content4.6.4 Glutamine Supplementation; 4.6.5 Culture Temperature; 4.7 Summary; References; 5: Isolation Of€pancreatic Islets From€nonhuman Primates; 5.1 Introduction; 5.2 Description Of€the€procedure And€key Aspects; 5.2.1 Laboratory Set Up; 5.2.1.1 Procedures Prior To€the€day Of€islet Isolation; 5.2.1.2 Procedures On€the€day Of€islet Isolation; Solutions And€instruments; Preparation Of€digestive Enzymes; Pancreas Procurement; Collagenase Digestion And€dilution; Concentration And€purification Of€islets; Determination Of€islet Yield And€purity. Islet Assessment In€nhp Isletsin Vitro Tests; In Vivo Test; 5.3 Conclusions; References; 6: Necessities For€a€clinical Islet Program; 6.1 Introduction; 6.2 The Islet Processing Facility; 6.3 Clinical Islet Isolation Outcomes; 6.4 The Donor Organ; 6.4.1 Effect Of€donor Age; 6.4.2 Effect Of€bmi; 6.4.3 Effect Of€cold Ischaemic Time (cit); 6.5 Organ Retrieval; 6.6 Pancreas Preservation; 6.6.1 The Two-layer Method; 6.6.2 Machine Perfusion; 6.7 The Pancreas; 6.8 Concluding Remarks; References; 7: Clinical Islet Isolation. Miriam Ramírez-domínguez, Editor. Includes Bibliographical References And Index. Pancreatic islets make up the endocrine pancreas and they contain the only source of insulin in the body, beta cells. Hence, access to high quality preparations of pancreatic islets is fundamental for in vitro studies and to test pre-clinical applications in animal models in vivo. Access to healthy human islets is also crucial to improve transplantation procedures for diabetes. Given the susceptibility of pancreatic islets to the enzymatic digestion and mechanical stress required to obtain them, the isolation of islets is often considered as the delicate 2work of a craftsman3. This book, which is aimed at beginners and experts alike, is a survey of the current state-of{u2013}the-art in this field and it centres on the challenges, pitfalls and peculiarities of pancreatic islet isolation in the different species used in pre-clinical and clinical applications. It explores the similarities and differences between human islets and those from other relevant species (rodents, pigs and non-human primates), and how these influence islet isolation. The ultimate goal of this book is to improve the outcome of islet isolation and transplantation in pre-clinical and clinical applications Front Matter....Pages i-xii Historical Background of Pancreatic Islet Isolation....Pages 1-9 The Different Faces of the Pancreatic Islet....Pages 11-24 Isolation of Mouse Pancreatic Islets of Langerhans....Pages 25-34 Pancreatic Islets: Methods for Isolation and Purification of Juvenile and Adult Pig Islets....Pages 35-55 Isolation of Pancreatic Islets from Nonhuman Primates....Pages 57-66 Necessities for a Clinical Islet Program....Pages 67-88 Clinical Islet Isolation....Pages 89-122 Back Matter....Pages 123-123