Porcine Aorta Isolation: A Surgical Procedure to Excise the Thoracic Aorta From Miniature Pigs for Downstream Analysis

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.

1. Surgery and preparation for porcine aortic endothelial cell isolation

1. Disinfecting the operating room with UV light for 1 h and medical disinfectant 84 liquid (available chlorine content is 4.0% – 5.0%, Table of Materials) before performing surgery.
2. After the miniature pig received an intramuscular injection formed by 15 mg/kg Ketamine, 15 mg/kg Xylazine hydrochloride and 0.05 mg/kg Phenacetin hydrochloride (Volumes: 100μL/kg, Table of Materials) (Figure 1A), confirm the animal's anesthesia status with checking porcine painful stimulus (eg. pinching one ear or shaking one forelimb) and physiological index (eg. blood pressure, heart and respiratory rate).
   NOTE: Another half dose injection could be given to the pig if it shows the signs of waking up.
3. Cut the abdomen with a scalpel, expose the inferior vena cava and consequently inject heparin sodium (5,000 U, Table of Materials) into the inferior vena cava with 5 mL syringe.
4. Five minutes later, insert a catheter to the abdominal aorta to blood, cut the skin of chest and incise the diaphragm with a scalpel and subsequently cut the left ventricle off.
   NOTE: Make sure that pig is euthanized by checking aortic pulse after blooding and cutting the heart.
5. Excise the ribs with bone forceps (Table of Materials) and expose the heart and lungs. Find the aorta posterior to the heart and lungs and clamp the two ends. Wash the aorta once with pre-cooled washing buffer- 1x PBS solution (pH 7.4) with 1% (v/v) Penicillin/Streptomycin. (Figure 1B,C).
6. Cut out the aorta with scissors and keep the aorta clamped at each end. Excise excess tissue around the aorta with sterile forceps and scissors. Put the aorta into a 50 mL centrifuge tube, wash the aorta with pre-cooled washing buffer 3x (30 mL per wash), and transfer the aorta to the laboratory (Figure 1D).