Rodent Transplant Medicine

Description:

This book introduces transplantation in rodents as useful tools used in studying transplant immunobiology. Several solid organs (kidney, heart, liver) transplant models in rodents are described in this book. It can help surgical quality and save surgical time. The first part of the book provides a review of rodent transplant tolerance induction, the role of gender and body-weight in rodent transplantation, surgical instruments and organ preservation solutions. In the second part of the book, various organ-transplantation techniques in rodents are discussed in individual chapters. This book presents uniform surgical procedures in mouse and rats, which produce comparable data, efficiently enhancing the translational research from bench to non-human primates and beyond. It will be of great value to transplant researchers, research fellows and clinicians in many surgical specialties.

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Foreword

With the rapid development of microsurgery and molecular biology, rodent transplant models have become increasingly popular. Transplantation in rodents is the most useful in studying transplant immunobiology, in which more immunological tools can be utilized. However, the highly demanding techniques not only hamper the wide use of rodent transplant models but also affect the production of reliable, comparable, reproducible data. Many surgical modifications can de facto cause a significant variation of allograft survival. Systematic description of commonly used transplant models of solid organs (kidney, heart, liver) in rodents may improve surgical quality and save surgical time as a consequence, which will reduce and refine animal use. Adoption of uniform surgical procedures can produce comparable data efficiently enhancing the research translation from bench to nonhuman primate, even bedside. Furthermore, master of microsurgical techniques will benefit trainees in many surgical specialties and surgical research as well.

Indeed, the innovation of technologies can interplay with the progress of science. The refinement of microsurgical techniques has significantly promoted the development of transplant immunology because it has made the feasibility of rodent transplant models, in which a large amount of monoclonal antibodies, reagents, knockout/ knockin animals, and other immunological tools can be utilized, but not for other laboratory species such as rabbits, dogs, swine, and nonhuman primates.

Currently, commercialized knockout rats have become available for research. Compared with the mouse, the rat is much more representative and indicative of human physiology, metabolism, and disease. The rat is advantageous in its larger size and physiological, cognitive, and breeding characteristics. Rat physiology is more suitable for studying human pharmacokinetics, toxicity, and immunology. Furthermore, more surgical procedures can be performed, and blood collection can be done simply and frequently in the larger animal.

As described above, establishment of mouse and rat transplant models is of great value in application for both scientific research and improvement of microsurgical techniques. Rodent Transplant Medicine by Dr. Weihua Gong et al. is directed at shedding light on the successful establishment of rodent transplant models and understanding rodent transplant biology. The chapters written by German, American, and Chinese experts are accomplished by clear illustrations. It will help the reader better grasp the microsurgical techniques of rodent transplant models.

In addition, the challenge of translational medicine from animal models to human situations exists in practice, which requires study models closely mimicking clinical scenarios. Other influencing factors including donor factors (prolonged warm/cold ischemic time, hypertension, diabetes, and brain death) and recipient factors (coinfections, immunological memory) should be taken into account. These aspects might be further discussed for the future in the forward.

Table of contents :

Foreword
Acknowledgments
Contents
Abbreviations
Chapter 1: Rodent Transplantation Medicine
1.1 Introduction
1.2 Advantages of Microsurgical Training in the Laboratory
1.3 Availability of Knockout Rat for Transplant Research
1.4 Tolerability of Different Transplant Organs
1.5 Vascularized Versus Nonvascularized Grafts
1.6 Comparison Between Mouse and Rat Transplant Models
1.7 Different Combinations of Rodents for Transplant Models
References
Chapter 2: T-Cell Costimulatory Molecules
2.1 The Ig Superfamily Costimulatory Molecules
2.1.1 CD28/CTLA-4/B7 Pathways
2.1.2 ICOS/B7h Pathway
2.1.3 PD-1/PD-L1/PD-L2 Pathways
2.1.4 BTLA and CD160 Pathways
2.1.4.1 The TNF-TNFR Superfamily Costimulatory Molecules
2.1.5 CD40/CD154 Pathway
2.1.6 OX40/OX40L Pathway
2.1.6.1 4-1BB/4-1BBL Pathway
2.1.7 CD27/CD70 Pathway
2.1.8 CD30/CD30L Pathway
2.1.9 GITR/GITRL Pathway
2.1.10 HVEM/LIGHT Pathway
2.2 The TIM Family Costimulatory Molecules
2.3 Costimulatory Control of T Regulatory Cells
2.4 Conclusions
References
Chapter 3: Age in Rodent Organ Transplantation
3.1 Introduction of Clinical Observation on the Role of Age in Transplantation
3.2 Experimental Studies for Underlying Mechanism
3.3 Relationship Between Age and Alloimmune Responses
3.4 Role of Senescence of Graft
3.5 Synergistic Effect of Donor Age and Prolonged Cold/Warm Ischemia Time
3.6 Conclusive Remarks
References
Chapter 4: Role of Gender in Animal Transplantation
4.1 Introduction
4.2 Underlying Mechanism of the Effect of Gender Impairing on Transplant Rejection
4.3 Conclusive Remarks
References
Chapter 5: Role of Body Weight/Functioning Mass of Graft in Rodent Transplantation
5.1 The Difference of Growth Curves Between Rat and Mouse
5.2 Clinical Observation of a Significance of Body Mass Index (BMI)
5.3 Experimental Findings on the Role of Donor Organ Weight or Body Weight
5.4 Underlying Mechanisms for the Effect of Body Weight/Functioning Mass of Graft
References
Chapter 6: Illustration of Laboratory Facilities and Microsurgical Instruments
6.1 Introduction
6.2 Animal Training Program
6.3 Certificate of Animal Training Program
6.4 Anesthetics
6.5 Microsurgical Instruments
6.5.1 Operating Microscope
6.5.2 Bipolar Coagulator
6.5.3 Low-Temperature Cautery
6.5.4 Rodent Operating Board
6.5.5 Dual Clasping Stainless Steel Sterilization Container
6.5.6 Angiocatheter and Bent 30-Gauge Needle
6.5.7 Retractors
6.5.8 Microvascular Clamps
6.5.9 Curved Needle Holder
6.5.10 Microclip Vessel Applier with Lock
6.5.11 Sterile Swab (Cotton Fioc) and Gauze
6.5.12 Microsurgical Scissor and Microdissecting Forceps Fine Point
6.5.13 Needle and Sutures
6.5.14 6-0 Silk
6.5.15 Surgifoam
6.5.16 Autoclip Wound Clip Applier (9 mm)
6.5.17 Circulating Water Heating Pad and Pump
6.5.18 Metabolic Cage
References
Chapter 7: Organ Preservation
7.1 Rat Kidney Transplantation
7.2 Rat Liver Transplantation
7.3 Mouse Heart Transplantation
7.4 Mouse Kidney Transplantation
7.5 Mouse Liver Transplantation
References
Chapter 8: Rat Kidney Transplantation
8.1 Rat Kidney Transplant Models
8.2 Animals
8.3 Anesthesia
8.4 Donor Surgery
8.5 Recipient Surgery
8.6 Vascular Anastomosis
8.7 Reperfusion
8.8 Ureter Anastomosis
8.9 Postoperative Care
8.10 Graft Assessment
8.11 Discussion
References
Chapter 9: Rat Orthotopic Liver Transplantation
9.1 Animal
9.2 Anesthetization
9.3 Donor Procedure
9.4 Donor Back Table Procedure
9.5 Recipient Procedure
9.6 Discussion
9.7 Selection for Recipient and Donor
9.8 Narcotic Control and Choice
9.9 Donor Liver Surgery and Back Table Procedure
9.10 Receptors Surgery
References
Chapter 10: Mouse Skin Transplantation
10.1 Special Surgical Tool
10.1.1 Preservation Solution
10.2 Surgical Procedure
10.2.1 Donor Surgery
10.2.1.1 Trunk Skin Surgery
10.2.1.2 Tail Skin Procurement
10.2.1.3 Ear Skin Surgery
10.2.2 Recipient Surgery
10.2.2.1 Suture Technique
10.2.2.2 Tissue Adhesive Technique (TAT)
10.3 Dressing After Surgery
10.4 Disbandagement
10.5 Observation of Graft Survival
10.6 Histological Evaluation of Skin Graft
10.7 Analgesic
10.8 Admission of Graft for Analysis
10.9 Discussion
References
Chapter 11: Mouse Heterotopic Abdominal Heart Transplant Model
11.1 Introduction
11.2 Mice
11.3 Surgical Instruments
11.4 Anesthetic
11.5 Storage Solution for Donor Organ
11.6 Mouse Abdominal Heart Transplant Model
11.6.1 Donor Heart Preparation
11.7 Recipient Surgery
11.8 Arteriotomy and Venotomy
11.9 Vascular Reconstruction
11.10 Postoperative Care
11.11 Postoperative Analgesia
11.12 Graft Assessment
11.13 Admission Standard of Transplanted Recipients
11.14 Discussion
References
Chapter 12: Mouse Heterotopic Cervical Heart Transplant Model
12.1 Introduction
12.2 Donor Surgery
12.3 Recipient Operation
12.4 Discussion
12.5 Mouse Cervical Heart Transplant Model with Cuff Technique
12.5.1 Materials and Methods
12.5.1.1 Animal
12.5.1.2 Donor Procedure
12.5.1.3 Recipient Procedure
12.5.2 Discussion
References
Chapter 13: Mouse Kidney Transplantation
13.1 Animals
13.2 Special Surgical Instruments
13.3 Preparation of the Surgeon
13.4 Anesthesia
13.5 Donor Surgery
13.5.1 Cold Ischemia
13.5.2 Warm Ischemia
13.6 Recipient Surgery
13.6.1 Urethral Reconstruction
13.7 Postoperative Care
13.7.1 Body Temperature
13.7.2 Antibiotics
13.7.3 Analgesic
13.8 Nephrectomy for the Recipient’s Native Kidney
13.9 Admission Standard of Transplanted Recipients
13.10 Graft Assessment
13.11 Histological Analysis
13.12 Discussion
References
Chapter 14: Mouse Liver Transplantation
14.1 Surgical Procedure
14.2 Donor
14.3 Back Table
14.4 Recipient
14.5 Tips and Pitfalls
14.6 Complications and Rejection
14.7 Arterialization
References
Chapter 15: Retinal Stem Cell Transplantation for Rat
15.1 Introduction
15.2 Animal
15.3 Methods
References
Chapter 16: Islet Transplantation in Mice
16.1 Establishment of Mice Models of Diabetes Mellitus
16.1.1 Main Reagents and Preparation Methods
16.1.2 Establishment of Mice Models
16.2 Isolation of Mouse Islets
16.2.1 Main Reagents and Preparation Methods
16.2.2 Isolation of Mouse Islets
16.2.3 Purification of Mouse Islets
16.2.4 Identification of Islet Cells
16.2.5 Functional Testing of Islet Cells
16.3 Allogeneic Islet Cell Transplantation in Mice
16.3.1 Transplantation of Islet Cells Under the Renal Capsule in Mice
16.3.2 Blood Glucose Monitoring After Islet Transplantation
16.3.3 Statistical Analysis
16.4 Results
16.5 Discussions
References
Chapter 17: Mouse Hematopoietic Stem Cell Transplantation
17.1 Recipients and Donors
17.1.1 Recipient Mice
17.1.2 Donors
17.2 Conditioning Recipient Mice
17.2.1 Conditioning with TBI
17.2.2 Conditioning with Non-TBI Regimen
17.3 Harvesting HSCs from Donors
17.3.1 Harvesting HSCs from Adult Mice
17.3.2 Harvesting HSCs from Embryonic Mice
17.3.3 Harvesting HSCs from the Human Body
17.4 Route for HSC Transplantation
17.5 Assessment After HSC Transplantation
References
Appendix
Record of Experiments
Treatment
Animal Surgical Record
Daily Clinic Observation
Daily Clinic Observation
Daily Clinic Observation
Euthanasia Record
Index