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The present paper describes a modified technique for heterotopic vascularized cardiac transplantation with updated aseptic technique, analgesia, and anesthesia.
The development of experimental models of cardiac transplantation in animals has contributed to many advances in the fields of immunology and solid organ transplantation. While the heterotopic vascularized murine cardiac transplantation model was initially utilized in studies of graft rejection using combinations of mismatched inbred mouse strains, access to genetically modified strains and therapeutic modalities can provide powerful new preclinical insights. Fundamentally, the surgical methodology for this technique has not changed since its development, especially with respect to important factors such as aseptic technique, anesthesia, and analgesia, which make material impacts on postsurgical morbidity and mortality. Additionally, improvements in perioperative management are expected to provide improvements in both animal welfare and experimental outcomes. This paper reports upon a protocol developed in collaboration with a subject matter expert in veterinary anesthesia and describes the surgical technique with an emphasis on perioperative management. Additionally, we discuss the implications of these refinements and provide details on troubleshooting critical surgical steps for this procedure.
We owe much of our understanding of immunology and transplantation to research based on experimental models of solid organ transplantation using animal subjects. Since the first description of vascularized cardiac transplantation in mammals1, such models have contributed to knowledge in wide-ranging domains, including the therapeutic application of hypothermia2, the benefits of using specialized sutures3, and techniques for total lung and heart homotransplantations4. The development of cardiac transplantation models in rats5,
This research was performed in accordance with the Code of Practice for the Care and Use of Animals for Scientific Purposes15 and approved under Animal Ethics Protocols RA/3/100/1568 and AE173 (The University of Western Australia Animal Ethics Committee and The Harry Perkins Institute of Medical Research Animal Ethics Committee, respectively). See the Table of Materials for details regarding all materials, instruments, and animals used in this protocol.
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To determine the effectiveness of the surgical technique in promoting good outcomes of wound healing and mouse recovery, early experiments in the laboratory determined the survival characteristics of a range of heart grafts of variable immunogenicity to the recipient. These included congenic (n = 5) and syngeneic (n = 5) grafts, which share the same major histocompatibility complex (MHC) markers as the recipient, and major mismatch grafts (n = 9), in which the graft and the recipient have different MHC markers. We used d.......
The murine orthotopic heart transplant model is a robust preclinical model used primarily to investigate the effects of MHC mismatch on the level and nature of immunological rejection and, more recently, the effect of transplantation on the retention of graft tissue-resident immunity16. While initially closely following the Corry et al.7 protocol, we have refined the protocol to incorporate best-practice standards of aseptic technique, analgesia, and anesthesia. The updatin.......
The authors would like to acknowledge the superb efforts of the animal care staff of the University of Western Australia and of the Harry Perkins Institute of Medical Research, whose dedication and expertise contributed to the feasibility and success of these surgeries.
....Name | Company | Catalog Number | Comments |
2030 Rycroft irrigating cannula 30 G | McFarlane | 56005HU | |
Braided surgical silk 7-0 | |||
Bulldog clamp curved - 35 mm | Roboz | RS-7441-5 | |
Bupivacaine 0.25%Â | |||
Buprenorphine | |||
Castroviejo needle holder catch curved -Â 145 mm | Haag-Streit | 11.62.15 | |
Chlorhexidine 5% solution | Ebos | JJ61371 | |
Cotton-tipped applicator - 7.5 cm | Dove | SN109510 | |
Ethanol 70% solution | Ebos | WH130192EE | |
Gauze 5 x 5 cm white | Aero | AGS50 | |
Gelfoam 80 mm x 125 mm | Pfizer | 7481D | |
Hair clipper | Wahl | 9860L | |
Heparin 1,000 IU in 1 mL | |||
Iris SuperCut scissors straight - 11.5 cm  | Inka Surgical | 11550.11 | |
Isoflurane vaporiser | Darvall | 9176 | |
Micro bulldog clamp - 3.7 cm | Greman | 14119-G | |
Micro scissors curved 105 mm | |||
Micropore plain paper surgical tape - 2.5 cm wide | Ebos | 7810L | |
Microsurgical scissors - curved tip | |||
Monofilament polyprolene suture - 5/0 | Surgipro | P-205-X | |
Myweigh i101 Precision Scale 100 g x 0.005 g | Myweigh | Kit00053 | |
Needle - 30 G x 0.5 inch | BD | BD304000 | |
Needleholder 15 cm curved "super fine" | Surgical Specialists | ST-B-15-8.2 | |
Nylene 10/0 x 15 cm on 3.8 mm 3/8 circle round bodied taper (diam 0.07mm) CV300 | |||
Round body suture forceps curved 0.3 mm 120 mm | B. Braun | FD281R | |
Round body suture forceps straight 0.3 mm 120 mm | B. Braun | FD280R | |
Round handled vannas spring scissors-str/12.5 cm | 15400-12 | ||
Spring scissors-Cvd Sm blades | 15001-08 | ||
Stevens scissors blunt straight 110 mm | |||
Surgical backboard | Rigid laminated cardboard. 15 x 15 cm | ||
Surgical drapes | Cut into two sizes. 25 cm x 25 cm, and 25 cm x 40 cm | ||
Surgical microscope | |||
Syringe - 1 mL | BD | 592696 | |
Syringe - 3 mL | Leica | M651 | |
Toothed forceps | BD | 309657 | |
University of Wisconsin Solution | |||
Warming pad | Far infrared warming pad 20 x 25 cm | ||
Westcott spring scissors | |||
Yasargil clip applier bayonet | Aesculap | FE582K | |
Yasargil titanium clip perm 6.6 mm | Aesculap | A19FT222T | |
Mouse usage | |||
Strain/SEX/Weight | Donor | Recipent | |
BALB/c, female, 19-23 g | 7 | 21 | |
C57BL/6, female, 17-20 g | 7 | 0 | |
CD45.1 BALB/c, female, 17-21 g | 5 | 0 |
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