Both the heterotopic and Orthotopic tracheal transplantations begin with the removal of the donor trachea for the heterotopic transplantation. An abdominal incision is made in the recipient and the intestines are placed on a glove. The donor trachea is placed into the center of the greater omentum and fixed by two stitches for the orthotopic transplantation.
After a cervical incision is made in the recipient, the trachea is freed of surrounding tissue and cut. Then the donor trachea is inserted between the two ends of the recipient trachea and the trachea are stitched together. Hi, I'm Sonya Refa.
I'm professor in Immunology and cardiac surgery in the Department of Cardiovascular Surgery at the University Heart Center in Hamburg. My laboratory is focusing on transplant immunology and stem cell immunobiology. Today we will introduce you to Moon Auto topic and topic transplantation model of the twig here.
Hello, I'm from the TSI lab. We are using the also topic and the heterotopic trachea transplantation mouse models to study the mechanisms behind the development of observative airway diseases. And I'm Carrie Wan, a student at Stanford University and a summer fellow in Drer's TSI Laboratory.
So let's get started. Before beginning this procedure shaved the cervical hair and disinfect the area using beta isod. Confirm that the donor mouse is completely anesthetized by a lack of the toe pinch reflex.
Under microscopic view, perform a midline cervical incision from the level of the larynx to the sternum. Remove the subcutaneous fat as well as the strap muscles to get a clear view of the trachea. Dissect the trachea from any surrounding tissues such as the esophagus, nerves, arteries, and connective tissue.
Remove the whole trachea from the larynx to the bifurcation. Flush the transplant trachea with cold saline and store the graft at four degrees Celsius. After confirming a lack of the toe pinch reflex in the anesthetized recipient mouse, shave the abdominal hair in a wide margin around the incision site and disinfect the area.
Using beta is Sedona and alcohol. Perform a median laparotomy and place the intestine into a moisture glove. Spread the greater omentum.
Carefully place the graft into the center and fixate it with two eight oh proline sutures. Fully cover the transplant with the greater omentum and fix the graft with a single eight o prolene suture. Relocate the intestines back into the abdomen.
Close up the incision in two layers, the abdominal wall and skin layer with continuous pattern seven O proline sutures. After confirming that the recipient mouse is fully anesthetized using IP applicated injection, anesthesia, shave and disinfect the cervical area as previously shown, perform a midline cervical incision from the level of the larynx to the sternum. Divide the thymus and strap muscles to visualize the entire laryn tracheal complex.
Carefully dissect the trachea from the surrounding tissues. Taking care to preserve the recurrent laryngeal nerves. Spread the corvus and divide the trachea.
Three rings coddle from the cricoid. The animal will maintain physiologic respiration via the tracheostomy. Ensure clean tracheal edges in the recipient as well as in the graft by dissecting the tracheas under microscopic view.
The graft is interposed between the recipient tracheal defects and oriented to maintain anatomic polarity using eight o proline sutures. Anastomosis the donor graft with the distal or mediastinal trachea. The posterior aspect of the anastomosis is performed in continuous running fashion.
The anterior aspect is then completed using interrupted sutures. Then remove any secretions from the airway. The proximal anastomosis is then completed in the same way as the distal one.
Perform the posterior aspect in continuous running fashion and the anterior aspect with interrupted sutures. Ensure integrity of the airway and adequate spontaneous breathing. Relocate the strap muscles and close the skin layer using six oh continuous pattern Vicryl sutures when the heterotopic transplanted trachea is recovered after 28 days and analyzed histologically by h and d staining 100%luminal obliteration is observed when the orthotopic transplanted trachea is recovered after 60 days and analyzed histologically by h and d staining, a maximal luminal obliteration of approximately 45%is achieved.
We have just shown you two different models to investigate the development of literative airway disease. The appropriate model should be chosen by the study hypothesis. Our lab favors the also topic one since the physiological airflow imitates the clinical setting.
So that's it. Thanks for watching and good luck with your experiments.
