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DOI: 10.3791/163-v
Significant breakthroughs in stem cell identification are continuously being made. To translate these discoveries, however, novel methods for cellular delivery must be devised. Here I report that the airways provide a safe route for stem cell transplantation to the lungs.
With major progress in the field of stem cell science, the rate limiting step in therapy may involve efficient tissue delivery. Tracheostomy is a surgical procedure on the neck to open a direct route to the lung, which allows the distribution of stem cells. Hi, my name is Yako Petter.
I'm from the Department of Pulmonary and Critical Care Medicine at Harvard Medical School. Today, I will show you a procedure that will allow researchers to access the lung, in particular, the left lobe in a safe and reliable manner. This method involves a tracheal incision, cattle insertion of a cannula into and along the tracheal lumen and injection of a stem cell mixture into the lung.
Well, let's get started. To determine the dose of anesthesia, we wave the mouse. We administer the anesthesia by interperitoneal injection Prior to surgical operations, a toe pinch is performed to show that the animal is fully anesthetized.
Although not necessary, we prefer to stabilize the mouse by constraining him corly between suture thread at incisors and caly by taping the tail. After sterilizing the surgical area using Betadine and an alcohol swab, we expose a trachea by making a 0.5 to one C centimeter midline incision in the ventral aspect of the neck. Once the incision has been made, we carefully separate the tissue from the trachea to give us direct access.
A tie is then inserted underneath the trachea cautiously using the tie, the trachea is gently pulled upward. A needle is then inserted into the trachea smoothly, yet rapidly until you feel slight resistance. Remove the needle, leaving the cannula in place and pipe it up to 50 microliters of cells into the catheter.
Immediately remove the tie and tube while removing the mouse from its constraints. If needed, gently hold the mouse upright by clinging his anterior paws and moving gently. Now you can close the wound with one suture or antibiotic glue through the incision.
After the operation, it is recommended to maintain mouse body temperature. We do so by placing the subject under a heat lamp while protecting his eyes from the glare. Next, as a postoperative analgesia, we administer buprenorphine every 12 hours for the first 24 hours.
Following this procedure, we just showed you how to inject stem cells and other agents into the lung via the trachea. So that's it. Good luck with your experiments.
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