A Mouse Aortic Interposition Model to Study Alloimmune-Induced Vascular Rejection

Abstract

Source: Enns, W., et al. Mouse Model of Alloimmune-induced Vascular Rejection and Transplant Arteriosclerosis. J. Vis. Exp. (2015).

This video demonstrates a method for generating a mouse aortic interposition model to study alloimmune-induced vascular rejection. After placing an anesthetized donor mouse in the supine position, the infrarenal abdominal aorta (IAA) is carefully isolated from surrounding structures. Branching vessels are ligated, and both ends of the IAA are clamped to prevent blood flow before resection. Subsequently, the donor IAA is implanted into an allogeneic recipient mouse, and blood flow is restored through anastomosis to investigate alloimmune-induced vascular rejection.

Protocol

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board. Use Balb/cYJ (H2d) donor mice and C57Bl/6 (H2b) recipient mice to examine allogeneic reactions. Mice are used for experiments between the ages of 8 to 12 weeks. Use either female or male mice. Syngeneic controls consist of aortic segments from C57Bl/6 donors into C57Bl/6 recipients. 1. Donor and …

Materials

C57BL/6J (H-2b)	Jackson Laboratories, Bar Harbour ME	Strain# 000664
Balb/cBYJ	Jackson Laboratories, Bar Harbour ME	Strain# 001026
Ketamine Hydrochloride Injection USP 100 mg/ mL	Ketalean	DIN 00612316
Xylazine Injection 20 mg/mL	Rompum	DIN 02169592
Ketoprofen Injection 100 mg/mL	Anafen	DIN 01938126
Butorphanol Tartrate injection 10 mg/mL	Torbugesic	DIN 008450000
Buprenorphine Injection 0.3 mg/mL	Reckitt Benckiser	B.N. 5241
Atipamezole hydrochloride sterile injectable solution	Antisedan	DIN 02237744
Heparin Sodium Injection, USP, 1000 units/mL	McKesson Distribution	DIN 02264315
Tears naturale ophthalmic ointment	Alcon	DIN 02082519
Stereomicroscope	Leica	M80
0.9% Sodium Chloride, sterile	Baxter Corporation
Lactated Ringer's solution, sterile	Baxter Corporation
0.9% Sodium Chloride Injection, sterile, 10 mL	Baxter Corporation
Alcohol Prep Pads	Loris
Povidone Iodine	Betadine
Chlorohexidine Gluconate 4% w/v	Germi-Stat
Black Polyamide Monofilament	AROSurgical Instruments	T4A10Q07
Suture, 10-0 suture, 70 microns	Corporation
Blue monofilament suture 5-0, P3 needle	Ethicon	8698G
1 ml Syringe	BD	REF 309659
10 ml Syringe	BD	REF 309604
1cc TB insulin syringe with 28G 1/2	BD	REF 309309
25G 7/8, hypodermic needle	BD	REF 305124
27G 1/2, hypodermic needle	BD	REF 305109
Colibri Retractor- 1.5cm spread 4cm	Fine Science Tools	17000-04
S&T CAF-4 Clip applying forceps, without lock	Fine Science Tools	00072-14
Supergrip forceps, S&T	Fine Science Tools	00632-11
Medical No.5 forceps	Fine Science Tools	11253-20
Lexer Baby Scissors	Fine Science Tools	14078-10
Micro Adson forceps serrated	Fine Science Tools	11018-12
Vannas-Tubingen microscissors	Fine Science Tools	15003-08
Micro clamps, b-1; 3.5mm x 1mm; 7mm length	Fine Science Tools	00396-01
Graefe-forceps, 10cm 1×2 teeth	Fine Science Tools	11054-10
Castroviejo with lock and tungsten jaws	Fine Science Tools	12565-14
Hot glass bead sterilizer	Inotech 250	IS-250 – Steri-250
Non-woven gauzes	Progene
Cotton Tipped Applicators	Puritan
Beard Trimmer	Wahl
Heating pad	Sunbeam